Anti-gram-positive bacterial methods and materials

ABSTRACT

The present invention relates to methods of treating gram-positive bacterial infections by administration of a BPI protein product alone, or in combination with an antibiotic. BPI protein product alone has a bactericidal or growth inhibitory effect on selected gram-positive organisms. BPI protein product also increases the susceptibility of gram-positive organisms to antibiotics and can even reverse resistance of gram-positive organisms to antibiotic.

This is a Continuation of U.S. application Ser. No. 08/372,783, filedJan. 13, 1995, now U.S. Pat. No. 5,578,572, which is acontinuation-in-part of U.S. patent application Ser. No. 08/274,299filed Jul. 11, 1994, abandoned, which is a continuation-in-part of U.S.patent application Ser. No. 08/209,762 filed Mar. 11, 1994, now U.S.Pat. No. 5,733,872, which is a continuation-in-part of U.S. patentapplication Ser. No. 08/183,222 filed Jan. 14, 1994, abandoned, all ofwhich are incorporated herein by reference.

BACKGROUND OF THE INVENTION

The present invention relates generally to methods and materials fortreating gram-positive bacterial infections by administration ofbactericidal/permeability-increasing (BPI) protein products alone, or incombination with antibiotic substances. Also disclosed are analogousmethods and materials involving use of lipopolysaccharide bindingprotein (LBP) derivatives.

BPI is a protein isolated from the granules of mammalianpolymorphonuclear leukocytes (PMNs or neutrophils), which are bloodcells essential in the defense against invading microorganisms. HumanBPI protein has been isolated from PMNs by acid extraction combined witheither ion exchange chromatography Elsbach, J. Biol. Chem., 254:11000(1979)! or E. coli affinity chromatography Weiss, et al., Blood, 69:652(1987)!. BPI obtained in such a manner is referred to herein as naturalBPI and has been shown to have potent bactericidal activity against abroad strum of gram-negative bacteria. The molecular weight of human BPIis approximately 55,000 daltons (55 kD). The amino acid sequence of theentire human BPI protein and the nucleic acid sequence of DNA encodingthe protein have been reported in FIG. 1 of Gray et al., J. Biol. Chem.,264:9505 (1989), incorporated herein by reference. The Gray et al. aminoacid sequence is set out in SEQ ID NO: 69 hereto.

BPI is a strongly cationic protein. The N-terminal half of BPI accountsfor the high net positive charge; the C-terminal half of the moleculehas a net charge of -3. Elsbach and Weiss (1981), supra.! A proteolyticN-terminal fragment of BPI having a molecular weight of about 25 kD hasan amphipathic character, containing alternating hydrophobic andhydrophilic regions. This N-terminal fragment of human BPI possesses theanti-bacterial efficacy of the naturally-derived 55 kD human BPIholoprotein. Ooi et al., J. Bio. Chem., 262: 14891-14894 (1987)!. Incontrast to the N-terminal portion, the C-terminal region of theisolated human BPI protein displays only slightly detectableanti-bacterial activity against gram-negative organisms. Ooi et al., J.Exp. Med., 174:649 (1991).! An N-terminal BPI fragment of approximately23 kD, referred to as "rBPI₂₃," has been produced by recombinant meansand also retains anti-bacterial activity against gram-negativeorganisms. Gazzano-Santoro et al., Infect. Immun. 60:4754-4761 (1992).

The bactericidal effect of BPI has been reported to be highly specificto gram-negative species, e.g., in Elsbach and Weiss, Inflammation:Basic Principles and Clinical Correlates, eds. Gallin et al., Chapter30, Raven Press, Ltd. (1992). BPI is commonly thought to be non-toxicfor other microorganisms, including yeast, and for higher eukaryoticcells. Elsbach and Weiss (1992), supra, reported that BPI exhibitsanti-bacterial activity towards a broad range of gram-negative bacteriaat concentrations as low as 10⁻⁸ to 10⁻⁹ M, but that 100- to 1,000-foldhigher concentrations of BPI were non-toxic to all of the gram-positivebacterial species, yeasts, and higher eukaryotic cells tested at thattime. It was also reported that BPI at a concentration of 10⁻⁶ M or 160μg/ml had no toxic effect, when tested at a pH of either 7.0 or 5.5, onthe gram-positive organisms Staphylococcus aureus (four strains),Staphylococcus epidermidis, Streptococcus faecalis, Bacillus subtilis,Micrococcus lysodeikticus, and Listeria monocytogenes. BPI at 10⁻⁶ Mreportedly had no toxic effect on the fungi Candida albicans and Candidaparapsilosis at pH 7.0 or 5.5, and was non-toxic to higher eukaryoticcells such as human, rabbit and sheep red blood cells and several humantumor cell lines. See also Elsbach and Weiss, Advances in InflammationResearch, ed. G. Weissmann, Vol. 2, pages 95-113 Raven Press (1981).This reported target cell specificity was believed to be the result ofthe strong attraction of BPI for lipopolysaccharide (LPS), which isunique to the outer membrane (or envelope) of gram-negative organisms.

The precise mechanism by which BPI kills gram-negative bacteria is notyet completely elucidated, but it is believed that BPI must first bindto the surface of the bacteria through electrostatic and hydrophobicinteractions between the cationic BPI protein and negatively chargedsites on LPS. LPS has been referred to as "endotoxin" because of thepotent inflammatory response that it stimulates, i.e., the release ofmediators by host inflammatory cells which may ultimately result inirreversible endotoxic shock. BPI binds to lipid A, reported to be themost toxic and most biologically active component of LPS.

In susceptible gram-negative bacteria, BPI binding is thought to disruptLPS structure, leading to activation of bacterial enzymes that degradephospholipids and peptidoglycans, altering the permeability of thecell's outer membrane, and initiating events that ultimately lead tocell death. Elsbach and Weiss (1992), supra!. BPI is thought to act intwo stages. The first is a sublethal stage that is characterized byimmediate growth arrest, permeabilization of the outer membrane andselective activation of bacterial enzymes that hydrolyze phospholipidsand peptidoglycans. Bacteria at this stage can be rescued by growth inserum albumin supplemented media Mannion et al., J. Clin. Invest.,85:853-860 (1990)!. The second stage, defined by growth inhibition thatcannot be reversed by serum albumin, occurs after prolonged exposure ofthe bacteria to BPI and is characterized by extensive physiologic andstructural changes, including apparent damage to the inner cytoplasmicmembrane.

Initial binding of BPI to LPS leads to organizational changes thatprobably result from binding to the anionic groups in the KDO region ofLPS, which normally stabilize the outer membrane through binding of Mg⁺⁺and Ca⁺⁺. Attachment of BPI to the outer membrane of gram-negativebacteria produces rapid permeabilization of the outer membrane tohydrophobic agents such as actinomycin D. Binding of BPI and subsequentgram-negative bacterial killing depends, at least in part, upon the LPSpolysaccharide chain length, with long O-chain bearing, "smooth"organisms being more resistant to BPI bactericidal effects than shortO-chain bearing, "rough" organisms Weiss et al., J. Clin. Invest. 65:619-628 (1980)!. This first stage of BPI action, permeabilization of thegram-negative outer envelope, is reversible upon dissociation of theBPI, a process requiring the presence of divalent cations and synthesisof new LPS Weiss et al., J. Immunol. 132: 3109-3115 (1984)!. Loss ofgram-negative bacterial viability, however, is not reversed by processeswhich restore the envelope integrity, suggesting that the bactericidalaction is mediated by additional lesions induced in the target organismand which may be situated at the cytoplasmic membrane (Mannion et al.,J. Clin. Invest. 86: 631-641 (1990)). Specific investigation of thispossibility has shown that on a molar basis BPI is at least asinhibitory of cytoplasmic membrane vesicle function as polymyxin B (In'tVeld et al., Infection and Immunity 56: 1203-1208 (1988)) but the exactmechanism as well as the relevance of such vesicles to studies of intactorganisms has not yet been elucidated.

BPI is also capable of neutralizing the endotoxic properties of LPS towhich it binds. Because of its bactericidal properties for gram-negativeorganisms and its ability to neutralize LPS, BPI can be utilized for thetreatment of mammals suffering from diseases caused by gram-negativebacteria, such as bacteremia or sepsis.

U.S. Pat. No. 5,198,541 discloses recombinant genes encoding and methodsfor expression of BPI proteins, including BPI holoprotein and fragmentsof BPI. It also describes the use of N-terminal fragments of BPI proteinfor co-treatment of gram-negative bacterial diseases with certainantibiotics, specifically penicillin, cephalosporins, rifampicin andactinomycin D.

Antibiotics are natural chemical substances of relatively low molecularweight produced by various species of microorganisms, such as bacteria(including Bacillus species), actinomycetes (including Streptomyces) andfungi, that inhibit growth of or destroy other microorganisms.Substances of similar structure and mode of action may be synthesizedchemically, or natural compounds may be modified to producesemi-synthetic antibiotics. These biosynthetic and semi-syntheticderivatives are also effective as antibiotics. The major classes ofantibiotics are (1) the β-lactams, including the penicillins,cephalosporins and monobactams; (2) the aminoglycosides, e.g.,gentamicin, tobramycin, netilmycin, and amikacin; (3) the tetracyclines;(4) the sulfonamides and trimethoprim; (5) the fluoroquinolones, e.g.,ciprofloxacin, norfloxacin, and ofloxacin; (6) vancomycin; (7) themacrolides, which include for example, erythromycin, azithromycin, andclarithromycin; and (8) other antibiotics, e.g., the polymyxins,chloramphenicol and the lincosamides.

Antibiotics accomplish their anti-bacterial effect through severalmechanisms of action which can be generally grouped as follows: (1)agents acting on the bacterial cell wall such as bacitracin, thecephalosporins, cycloserine, fosfomycin, the penicillins, ristocetin,and vancomycin; (2) agents affecting the cell membrane or exerting adetergent effect, such as colistin, novobiocin and polymyxins; (3)agents affecting cellular mechanisms of replication, informationtransfer, and protein synthesis by their effects on ribosomes, e.g., theaminoglycosides, the tetracyclines, chloramphenicol, clindamycin,cycloheximide, fucidin, lincomycin, puromycin, rifampicin, otherstreptomycins, and the macrolide antibiotics such as erythromycin andoleandomycin; (4) agents affecting nucleic acid metabolism, e.g., thefluoroquinolones, actinomycin, ethambutol, 5-fluorocytosine,griseofulvin, rifamycins; and (5) drugs affecting intermediarymetabolism, such as the sulfonamides, trimethoprim, and thetuberculostatic agents isoniazid and para-aminosalicylic acid. Someagents may have more than one primary mechanism of action, especially athigh concentrations. In addition, secondary changes in the structure ormetabolism of the bacterial cell often occur after the primary effect ofthe antimicrobial drug.

The penicillins have a characteristic double-ring system composed of aβ-lactam ring, which provides the antibacterial activity, and athiazolidene ring. The penicillins are differentiated by a single sidechain that is unique for each penicillin. The compounds are bactericidaland act by inhibiting bacterial transpeptidase, an enzyme involved insynthesis of the bacterial cell wall. Because of their mechanism ofaction, penicillins are generally active against growing, but notresting, cells. Penicillins, especially penicillin G, have largelygram-positive activity; the relative insensitivity of gram-negative rodsto penicillin G and several other penicillins is probably due to thepermeability barrier of the outer membrane of gram-negative bacteria.Ampicillin, carbenicillin, ticarcillin, and some other penicillins areactive against gram-negative bacteria because they can pass through thisouter membrane. Penicillins have relatively few adverse effects, themost important of which are the hypersensitivity (allergic) reactions.These compounds are widely distributed in the body, but do not entercells and do not usually accumulate in CSF.

Bacterial resistance to the penicillins is by production of the enzymeβ-lactamase, which catalyzes hydrolysis of the β-lactam ring. Thepercentage of bacteria resistant to penicillin has risen to about 80%.Several penicillins, including methicillin, oxacillin, cloxacillin,dicloxacillin and nafcillin, are not affected by the β-lactamase ofstaphylococci. These antibiotics are useful against mostβ-lactamase-producing species of Staphylococcus. However, a small numberof species are resistant even to these penicillins. Some penicillins,amoxicillin and ticarcillin, are marketed in combination with clavulanicacid, which is β-lactamase inhibitor that covalently binds to the enzymeand prevents it from hydrolyzing the antibiotics. Another inhibitor,sulbactam, is marketed in combination with ampicillin.

The cephalosporins are characterized by β-lactam ring, like thepenicillins, but have an adjacent dihydrothiazine ring instead of athiazolidene ring. For convenience, these compounds are generallyclassified by generations. The first generation includes cephalothin,cephapirin, cefazolin, cephalexin, cephradine and cefadroxil. Thesedrugs generally have excellent gram-positive activity except forenterococci and methicillin-resistant staphylococci, and have onlymodest gram-negative coverage. The second generation includescefamandole, cefoxitin, ceforanide, cefuroxime, cefuroxime axetil,cefaclor, cefonicid and cefotetan. This generation generally loses somegram-positive activity by weight and gains limited gram-negativecoverage. The third generation includes cefotaxime, moxalactam,ceftizoxime, ceftriaxone, cefoperazone and ceftazidime. These compoundsgenerally sacrifice further gram-positive activity by weight but gainsubstantial gram-negative coverage against Enterobacter and sometimesare active against Pseudomonas. The cephalosporins bind topenicillin-binding proteins with varying affinity. Once binding occurs,protein synthesis is inhibited. Cephalosporins are usually welltolerated; adverse effects include hypersensitivity reactions andgastrointestinal effects. Cephalosporins may interact with nephrotoxicdrugs, particularly aminoglycosides, to increase toxicity. Resistance tocephalosporins is mediated by several mechanisms, including productionof β-lactamase, although some strains that do not produce β-lactamaseare nevertheless resistant.

Imipenem is a N-formimidoyl derivative of the mold product thienamycin.It contains a β-lactam ring and somewhat resembles penicillin except fordifferences in the second ring. It has activity against bothgram-positive and gram-negative organisms and is resistant to mostβ-lactamases, although not those from Pseudomonas. It is marketed incombination with cilastin, a compound that inhibits inactivation ofimipenem in the kidney by renal dihydropeptidase I enzyme. Cilastinincreases the concentration of imipenem in urine, although not in blood.

Aztreonam is the first of a new group of antibiotics referred to as themonobactams. These agents have a β-lactam ring but lack the second ringcharacteristic of the penicillins and cephalosporins. It acts by bindingto penicillin-binding proteins, and produces long, filamentous bacterialshapes that eventually lyse. Aztreonam is active only against aerobicgram-negative bacteria, is susceptible to inactivation by someβ-lactamases, and has few adverse effects.

The aminoglycosides contain amino sugars linked to an aminocyclitol ringby glycosidic bonds. They have similar mechanisms of action andproperties, but differ somewhat in spectrum of action, toxicity, andsusceptibility to bacterial resistance. The compounds are bactericidal,with activity against both gram-positive and gram-negative organisms,and act by binding to proteins on the 30S ribosome of bacteria andinhibiting protein synthesis. The aminoglycosides also bind to isolatedLPS and have a very weak outer membrane permeabilizing effect. Taber etal., Microbiological Reviews 53: 439-457 (1987)); Kadurugamuwa et al.,Antimicrobial Agents and Chemotherapy, 37: 715-721 (1993); Vaara,Microbiological Reviews 56: 395-411 (1992)!. This class of antibioticsincludes amikacin, gentamicin, kanamycin, neomycin, netilmycin,paromomycin and tobramycin. The aminoglycosides are usually reserved formore serious infections because of severe adverse effects includingototoxicity and nephrotoxicity. There is a narrow therapeutic windowbetween the concentration required to produce a therapeutic effect,e.g., 8 μg/ml for gentamicin, and the concentration that produces atoxic effect, e.g., 12 μg/ml for gentamicin. Neomycin in particular ishighly toxic and is never administered parenterally.

Tetracyclines have a common four-ring structure and are closelycongeneric derivatives of the polycyclic naphthacenecarboxamide. Thecompounds are bacteriostatic, and inhibit protein synthesis by bindingto the 30S subunit of microbial ribosomes and interfering withattachment of aminoacyl tRNA. The compounds have some activity againstboth gram-positive and gram-negative bacteria; however, their use islimited because many species are now relatively resistant. Adverseeffects include gastrointestinal effects, hepatotoxicity with largedoses, and nephrotoxicity in some patients. This antibiotic classincludes tetracycline, chlortetracycline, demeclocycline, doxycycline,methacycline, minocycline and oxytetracycline.

The sulfonamides are derivatives of sulfanilamide, a compound similar instructure to para-aminobenzoic acid (PABA), which is an essentialprecursor for bacterial synthesis of folic acid. The compounds aregenerally bacteriostatic, and act by competitively inhibitingincorporation of PABA into tetrahydrofolic acid, which is a requiredcofactor in the synthesis of thymidines, purines and DNA. Sulfonamideshave a wide range of activity against gram-positive and gram-negativebacteria, but their usefulness has diminished with increasingly highprevalence of bacterial resistance. The sulfonamide class of antibioticsincludes sulfacytine, sulfadiazine, sulfamethizole, sulfisoxazole,sulfamethoxazole, sulfabenzamide and sulfacetamide. Adverse effectsinclude hypersensitivity reactions and occasional hematologicaltoxicity.

Trimethoprim is an inhibitor of the dihydrofolate reductase enzyme,which converts dihydrofolic to tetrahydrofolic acid, a required factorfor DNA synthesis. Adverse effects include gastrointestinal distress andrare hematological toxicity. Trimethoprim is also available incombination with sulfamethoxazole (also known as co-trimoxazole). Thecombination is usually bactericidal, although each agent singly isusually bacteriostatic. The combination is the drug of choice forSalmonella infections, some Shigella infections, E. coli traveler'sdiarrhea and Pneumocystis carinii pneumonia.

The fluoroquinolones and quinolones are derivatives of nalidixic acid, anaphthyridine derivative. These compounds are bactericidal, and impairDNA replication, transcription and repair by binding to the DNA andinterfering with DNA gyrase, an enzyme which catalyzes negativesupercoiling of DNA. The fluoroquinolones, which include norfloxacin,ciprofloxacin, and ofloxacin, and the quinolones, which includecinoxacin, have a broad spectrum of antimicrobial activity againstgram-negative and gram-positive organisms. These compounds distributewidely through extravascular tissue sites, have a long serum half-life,and present few adverse effects. Because of their effect on DNA, thedrugs are contraindicated in pregnant patients and in children whoseskeletal growth is incomplete.

Vancomycin is a glycopeptide, with a molecular weight of about 1500,produced by a fungus. It is primarily active against gram-positivebacteria. The drug inhibits one of the final steps in synthesis of thebacterial cell wall, and is thus effective only against growingorganisms. It is used to treat serious infections due to gram-positivecocci when penicillin G is not useful because of bacterial resistance orpatient allergies. Vancomycin has two major adverse effects, ototoxicityand nephrotoxicity. These toxicities can be potentiated by concurrentadministration of another drug with the same adverse effect, such as anaminoglycoside.

The macrolides are bacteriostatic and act by binding to the 50S subunitof 70S ribosomes, resulting in inhibition of protein synthesis. Theyhave a broad spectrum of activity against gram-positive andgram-negative bacteria and may be bacteriostatic or bactericidal,depending on the concentration achieved at sites of infection. Thecompounds distribute widely in body fluids. Adverse effects includegastrointestinal distress and rare hypersensitivity reactions. The mostcommon macrolide used is erythromycin, but the class includes othercompounds such as clarithromycin and azithromycin.

The polymyxins are a group of closely related antibiotic substancesproduced by strains of Bacillus polymyxa. These drugs, which arecationic detergents, are relatively simple, basic peptides withmolecular weights of about 1000. Their antimicrobial activity isrestricted to gram-negative bacteria. They interact strongly withphospholipids and act by penetrating into and disrupting the structureof cell membranes. Polymyxin B also binds to the lipid A portion ofendotoxin and neutralizes the toxic effects of this molecule. PolymyxinB has severe adverse effects, including nephrotoxicity andneurotoxicity, and should not be administered concurrently with othernephrotoxic or neurotoxic drugs. The drug thus has limited use as atherapeutic agent because of high systemic toxicity, but may be used forsevere infections, such as Pseudomonas aeruginosa meningitis, thatrespond poorly to other antibiotics.

Chloramphenicol inhibits protein synthesis by binding to the 50Sribosomal subunit and preventing binding of aminoacyl tRNA. It has afairly wide spectrum of antimicrobial activity, but is only reserved forserious infections, such as meningitis, typhus, typhoid fever, and RockyMountain spotted fever, because of its severe and fatal adversehematological effects. It is primarily bacteriostatic, although it maybe bactericidal to certain species.

Lincomycin and clindamycin are lincosamide antimicrobials. They consistof an amino acid linked to an amino sugar. Both inhibit proteinsynthesis by binding to the 50S ribosomal subunit. They compete witherythromycin and chloramphenicol for the same binding site but in anoverlapping fashion. They may be bacteriostatic or bactericidal,depending on relative concentration and susceptibility. Gastrointestinaldistress is the most common side effect. Other adverse reactions includecutaneous hypersensitivity, transient hematological abnormalities, andminor elevations of hepatic enzymes. Clindamycin is often the drug ofchoice for infections caused by anaerobic bacteria or mixedaerobic/anaerobic infections, and can also be used for susceptibleaerobic gram-positive cocci.

Some drugs, e.g. aminoglycosides, have a small therapeutic window. Forexample, 2 to 4 μg/ml of gentamicin or tobramycin may be required forinhibition of bacterial growth, but peak concentrations in plasma above6 to 10 μg/ml may result in ototoxicity or nephrotoxicity. These agentsare more difficult to administer because the ratio of toxic totherapeutic concentrations is very low. Antimicrobial agents that havetoxic effects on the kidneys and that are also eliminated primarily bythe kidneys, such as the aminoglycosides or vancomycin, requireparticular caution because reduced elimination can lead to increasedplasma concentrations, which in turn may cause increased toxicity. Dosesof antimicrobial agents that are eliminated by the kidneys must bereduced in patients with impaired renal function. Similarly, dosages ofdrugs that are metabolized or excreted by the liver, such aserythromycin, chloramphenicol, or clindamycin, must be reduced inpatients with decreased hepatic function.

Antibiotic resistance in bacteria is an increasingly troublesomeproblem. The accelerating development of antibiotic-resistant bacteria,intensified by the widespread use of antibiotics in farm animals andoverprescription of antibiotics by physicians, has been accompanied bydeclining research into new antibiotics with different modes of action.Science, 264: 360-374 (1994).! Antibiotic resistance, once acquired, canbe rapidly spread to other bacteria, including bacteria of a differentspecies. There are some species of bacteria that are resistant to allbut one antibiotic; it may be only a matter of time before theappearance of bacterial strains that are resistant to all antibiotics.

Bacteria acquire resistance to antibiotics through several mechanisms:(1) production of enzymes that destroy or inactivate the antibioticDavies, Science, 264:375-381 (1994)!; (2) synthesis of new or alteredtarget sites on or within the cell that are not recognized by theantibiotic Spratt, Science, 264:388-393 (1994)!; (3) low permeability toantibiotics, which can be reduced even further by altering cell wallproteins, thus restricting access of antibiotics to the bacterialcytoplasmic machinery; (4) reduced intracellular transport of the drug;and (5) increased removal of antibiotics from the cell viamembrane-associated pumps Nikaido, Science, 264:382-387 (1994)!.

The susceptibility of a bacterial species to an antibiotic is generallydetermined by two microbiological methods. A rapid but crude procedureuses commercially available filter paper disks that have beenimpregnated with a specific quantity of the antibiotic drug. These disksare placed on the surface of agar plates that have been streaked with aculture of the organism being tested, and the plates are observed forzones of growth inhibition. A more accurate technique, the brothdilution susceptibility test, involves preparing test tubes containingserial dilutions of the drug in liquid culture media, then inoculatingthe organism being tested into the tubes. The lowest concentration ofdrug that inhibits growth of the bacteria after a suitable period ofincubation is reported as the minimum inhibitory concentration.

The resistance or susceptibility of an organism to an antibiotic isdetermined on the basis of clinical outcome, i.e., whetheradministration of that antibiotic to a subject infected by that organismwill successfully cure the subject. While an organism may literally besusceptible to a high concentration of an antibiotic in vitro, theorganism may in fact be resistant to that antibiotic at physiologicallyrealistic concentrations. If the concentration of drug required toinhibit growth of or kill the organism is greater than the concentrationthat can safely be achieved without toxicity to the subject, themicroorganism is considered to be resistant to the antibiotic. Tofacilitate the identification of antibiotic resistance or susceptibilityusing in vitro test results, the National Committee for ClinicalLaboratory Standards (NCCLS) has formulated standards for antibioticsusceptibility that correlate clinical outcome to in vitrodeterminations of the minimum inhibitory concentration of antibiotic.

There continues to exist a need in the art for new antimicrobial, andespecially anti-gram-positive microbial, methods and materials. Productsand methods responsive to this need would ideally involve substantiallynon-toxic compounds available in large quantities by means of syntheticor recombinant methods. Ideal compounds would have bactericidal orbacteriostatic activity when administered or applied as the soleanti-microbial agent. Such compounds ideally would also be useful incombinative therapies with other anti-microbial agents, particularlywhere potentiating effects are provided.

SUMMARY OF THE INVENTION

The present invention provides methods and compositions for treatinggram-positive bacterial infections, including conditions associatedtherewith or resulting therefrom (for example, sepsis or bacteremia), ina subject by administering a BPI protein product alone, or concurrentlywith an antibiotic. The invention is based upon the discovery that BPIprotein products surprisingly have direct bactericidal and growthinhibitory effects on some gram-positive organisms, and that BPI proteinproducts unexpectedly have the ability to increase the antibioticsusceptibility of gram-positive bacteria, including the ability toreverse in many instances the antibiotic resistance of gram-positivebacteria. The invention is also based upon the finding that BPI proteinproducts and antibiotics provide additive and synergisticbactericidal/growth inhibitory effects when administered concurrently.

According to one aspect of the invention, a method is provided oftreating a gram-positive bacterial infection comprising the step ofadministering to a subject suffering from a gram-positive bacterialinfection a BPI protein product in an amount sufficient formonotherapeutic effectiveness. This method may be practiced when anyBPI-susceptible gram-positive bacterial species is involved in theinfection.

A second aspect of the invention provides a method of treatinggram-positive bacterial infection by concurrently administering to asubject suffering from a gram-positive bacterial infection a BPI proteinproduct in an amount sufficient for combinative therapeuticeffectiveness and one or more antibiotics in amounts sufficient forcombinative therapeutic effectiveness. This method is effective even forgram-positive organisms that are not susceptible to the directbactericidal/growth inhibitory effects of BPI.

For concurrent administration with antibiotics, the BPI protein productmay be administered in an amount effective to increase the antibioticsusceptibility of a gram-positive bacterial species involved in theinfection, or to potentiate the effects of the antibiotic. The BPIprotein product may also be administered in an amount effective toreverse the antibiotic resistance of a gram-positive bacterial speciesinvolved in the infection. The BPI protein product and the antibioticsmay each be administered in amounts that would be sufficient formonotherapeutic effectiveness upon administration alone or may beadministered in less than monotherapeutic amounts.

Another aspect of the invention provides a method of treatinggram-positive bacterial infection by concurrently administering to asubject suffering from a gram-positive bacterial infection a BPI proteinproduct and one or more antibiotics, in synergistically effectiveamounts.

In addition, the invention provides a method of killing or inhibitinggrowth of gram-positive bacteria comprising contacting the bacteria witha BPI protein product alone, or in combination with anotherantibacterial agent. This method can be practiced in vivo or in avariety of in vitro uses such as use in food preparation, todecontaminate fluids and surfaces or to sterilize surgical and othermedical equipment and implantable devices, including prosthetic joints.These methods can correspondingly be used for in situ sterilization ofindwelling invasive devices such as intravenous lines and catheters,which are often foci of infection, or for sterilization of in vitrotissue colture media.

The invention further provides a method of killing or inhibiting growthof gram-positive bacteria with disrupted cell walls, including L-phasevariants, by contacting such bacteria with a BPI protein product alone,or in conjunction with another antibacterial agent. This method can alsobe practiced on organisms that lack cell walls, such as Mycoplasma orUreaplasma.

The invention further provides the use of BPI protein product for themanufacture of a medicament for treatment of gram-positive bacterialinfections, and the use of a BPI protein product in combination with anantibiotic for manufacture of a medicament for treatment ofgram-positive bacterial infections.

Numerous additional aspects and advantages of the invention will becomeapparent to those skilled in the art upon consideration of the followingdetailed description of the invention which describes presentlypreferred embodiments thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 relates to plate assays of the bactericidal effect of rBPI₂₃ onBacillus subtilis.

FIG. 2 shows results from radial diffusion assays of the growthinhibitory effect of rBPI₂₁ on Staphylococcus aureus with or withoutpenicillin.

FIG. 3 depicts results from radial diffusion assays of the effect ofrBPI₂₁ on penicillin-treated S. aureus and L-phase variants.

FIG. 4 shows the effect of various BPI protein products onpenicillin-treated S. aureus in a radial diffusion assay.

FIGS. 5, 6, 7 and 8 show the effect of various BPI protein products,including BPI-derived peptides, on the S. aureus bacterial form andL-phase variant in a radial diffusion assay.

FIG. 9 relates to the effect of CaCl₂ concentration on the growth of S.aureus L-phase variant in broth.

FIG. 10 depicts results from a broth growth inhibition assay of theeffect of rBPI₂₁ on S. aureus L-phase variant.

FIGS. 11, 12 and 13 shows results from broth growth inhibition assays ofthe effect of rBPI₂₁ on S. aureus L-phase variant, at CaCl₂concentrations of 2.5 mM, 5 mM and 10 mM, respectively.

FIG. 14 shows the effect of rBPI₂₁ in an E. coli broth growth inhibitionassay.

FIG. 15 displays the effect of a variety of BPI protein products on S.pneumoniae L-forms in a radial diffusion assay.

FIGS. 16 and 17 show the effect of BPI protein products, includingBPI-derived peptides, on the S. pneumoniae bacterial form and L-phasevariant in a radial diffusion assay.

FIGS. 18 and 19 show the effect of BPI protein products, includingBPI-derived peptides, on the S. pyogenes bacterial form and L-phasevariant in a radial diffusion assay.

FIG. 20 shows the effect of a variety of BPI protein products on E.faecalis L-forms in a radial diffusion assay.

FIGS. 21 and 22 show the effect of BPI protein products, includingBPI-derived peptides, on the mycoplasma Acholeplasma laidlawii in aradial diffusion assay.

FIGS. 23, 24 and 25 display the effect of BPI protein products and LBPderivatives on S. aureus L-forms and S. pneumoniae L-forms in radialdiffusion assays.

FIGS. 26 through 32 displays rBPI₂₁ potentiation of the earlybactericidal effect of various antibiotics on S. pneumoniae, S. aureusand E. faecalis.

DETAILED DESCRIPTION

The present invention relates to methods and materials for treatingsubjects suffering from gram-positive bacterial infections."Gram-positive bacterial infection," as used herein, encompassesconditions associated with or resulting from gram-positive bacterialinfection (e.g., sequelae). These conditions include gram-positivesepsis and one or more of the conditions associated therewith, includingbacteremia, fever, hypotension, shock, metabolic acidosis, disseminatedintravascular coagulation and related clotting disorders, anemia,thrombocytopenia, leukopenia, adult respiratory distress syndrome andrelated pulmonary disorders, renal failure and related renal disorders,hepatobiliary disease and central nervous system disorders. Theseconditions also include translocation of gram-negative bacteria from theintestines and concomitant release of endotoxin. Gram-positive bacteriainclude bacteria from the following species: Staphylococcus,Streptococcus, Micrococcus, Peptococcus, Peptostreptococcus,Enterococcus, Bacillus, Clostridium, Lactobacillus, Listeria,Erysipelothrix, Propionibacterium, Eubacterium, and Corynebacterium.

A variety of gram-positive organisms are capable of causing sepsis. Themost common organisms involved in sepsis are Staphylococcus aureus,Streptoccocus pneumoniae, coagulase-negative staphylococci,beta-hemolytic streptococci, and enterococci, but any gram-positiveorganism may be involved. Bone, J. Critical Care, 8:51-59 (1993).!

According to one aspect of the invention, BPI protein product alone, inan amount sufficient for monotherapeutic effectiveness, may beadministered to a subject suffering from infection involving aBPI-susceptible gram-positive bacteria. When used to describeadministration of BPI protein product alone, the term "amount sufficientfor monotherapeutic effectiveness" means an amount of BPI proteinproduct that provides bactericidal or growth inhibitory effects whenadministered as a monotherapy. The invention utilizes any of the largevariety of BPI protein products known to the art including natural BPIprotein, recombinant BPI protein, BPI fragments, BPI analogs, BPIvariants, and BPI-derived peptides.

This aspect of the invention is based on the discovery that BPI proteinproducts have direct bactericidal or growth inhibitory activity againstsome gram-positive organisms. BPI protein products are also shown hereinto have direct bactericidal or growth inhibitory effects on L-phasevariants of a variety of gram-positive organisms; these L-phase variantslack the cell walls of the normal bacterial form. BPI protein productsare also expected to exert direct bactericidal/growth inhibitory effectson the cell wall-less Mycoplasma and Ureaplasma, organisms involvedclinically in respiratory and urogenital infections. Mycoplasma is alsoa major contaminant of in vitro tissue cultures.

According to a second aspect of the invention, a subject suffering froma gram-positive bacterial infection may be treated by concurrentadministation of a BPI protein product in an amount sufficient forcombinative therapeutic effectiveness and one or more antibiotics inamounts sufficient for combinative therapeutic effectiveness. Thisaspect of the invention contemplates concurrent administration of BPIprotein product with any antibiotic or combinations of antibiotics,including β-lactam antibiotics with and without β-lactamase inhibitors,aminoglycosides, tetracyclines, sulfonamides and trimethoprim,vancomycin, macrolides, fluoroquinolones and quinolones, polymyxins andother antibiotics.

This aspect of the invention is based on the discovery thatadministration of BPI protein products improves the therapeuticeffectiveness of antibiotics, e.g., by increasing the antibioticsusceptibility of gram-positive organisms to a reduced dosage ofantibiotics providing benefits in reduction of cost of antibiotictherapy and/or reduction of risk of toxic responses to antibiotics. BPIprotein products are shown herein to lower the minimum concentration ofantibiotics needed to inhibit in vitro growth of gram-positive organismsat 24 hours. In cases where BPI protein product did not affect growth at24 hours, BPI protein product was shown to potentiate the earlybactericidal effect of antibiotics in vitro at 0-7 hours. The BPIprotein products exert these effects even on gram-positive organismsthat are not susceptible to the direct bactericidal or growth inhibitoryeffects of BPI protein product alone.

This aspect of the invention is correlated to the additional discoverythat administration of a BPI protein product can effectively reverse theantibiotic resistance of a gram-positive organism. BPI protein productsare shown herein to reduce the minimum inhibitory concentration ofantibiotics from a level within the clinically resistant range to alevel within the clinically susceptible range. BPI protein products thuscan convert normally antibiotic-resistant organisms intoantibiotic-susceptible organisms.

According to this second aspect of the invention, the BPI proteinproduct and antibiotics are concurrently administered in amountssufficient for combinative therapeutic effectiveness. When used todescribe administration of a BPI protein product in conjunction with anantibiotic, the term "amount sufficient for combinative therapeuticeffectiveness" with respect to the BPI protein product means at least anamount effective to increase the susceptibility of the organism to theantibiotic, and the term "amount sufficient for combinative therapeuticeffectiveness" with respect to an antibiotic means at least an amount ofthe antibiotic that produces bactericidal or growth inhibitory effectswhen administered in conjunction with that amount of BPI proteinproduct. Either the BPI protein product or the antibiotic, or both, maybe administered in an amount below the level required formonotherapeutic effectiveness against a gram-positive bacterialinfection. The BPI protein product may be administered in an amountwhich is not sufficient for monotherapeutic effectiveness but whichprovides increased antibiotic susceptibility or potentiates the effectsof the antibiotic, or which reverses the resistance of the gram-positiveorganism to an antibiotic.

A further aspect of the invention relates to the discovery thatconcurrent administration of BPI protein products with antibioticsprovides synergistic bactericidal or growth inhibitory effects beyondthe individual bactericidal or growth inhibitory effects of the BPIprotein product or the antibiotic. Some methods for evaluating theeffect of antimicrobial combinations are described in Eliopoulos andMoellering In Antibiotics in Laboratory Medicine, 3rd ed. (Lorian, V.,Ed.) pp 432-492, Williams and Wilkins, Baltimore Md. (1991). There isgeneral agreement on qualitative definition of synergism: the combinedeffect of the drugs being examined is significantly greater than theexpected result based on independent effects of the drugs when usedseparately. In a checkerboard assay, the combination of BPI proteinproduct with antibiotics may be shown to result in a "synergistic"fractional inhibitory concentration index (FIC). The checkerboard methodis based on additivity, which assumes that the result observed withmultiple drugs is the sum of the separate effects of the drugs beingtested; according to this system a FIC of less than 0.5 is scored assynergy, 1 is scored as additive, and greater than 1 but less than 2 isscored as indifferent. In contrast, kinetic assays are based on the ideathat only one metabolic pathway at a time can be growth rate-limitingfor an organism; according to this system, the combined effect of drugsthat do not interact with one another (autonomous or indifferent) issimply the effect of the most active drug alone.

According to this aspect of the invention, "effective synergy" orpotentiation upon concurrent administration of BPI protein product withone or more antibiotics can be obtained in a number of ways. A BPIprotein product may convert an antibiotic-resistant organism into anantibiotic-susceptible organism or otherwise improve the antibioticsusceptibility of that organism. Conversely, a BPI-potentiatingantibiotic, such as an antibiotic that acts on the cell wall or cellmembrane of bacteria, may convert a BPI-resistant organism into aBPI-susceptible organism. Alternatively, the BPI protein product andantibiotic may both co-potentiate the other agent's activity. The BPIprotein product and antibiotic may have a therapeutic effect when bothare given in doses below the amounts sufficient for monotherapeuticeffectiveness.

Either the BPI protein product or the antibiotics may be administeredsystemically or topically. Systemic routes of administration includeoral, intravenous, intramuscular or subcutaneous injection (includinginto a depot for long-term release), intraocular or retrobulbar,intrathecal, intraperitoneal (e.g. by intraperitoneal lavage),transpulmonary using aerosolized or nebulized drug, or transdermal.Topical routes include administration in the form of salves, ophthalmicdrops, ear drops, or irrigation fluids (for, e.g., irrigation ofwounds).

"Concurrent administration," or co-treatment, as used herein includesadministration of the agents, in conjunction or combination, together,or before or after each other. The BPI protein product and antibioticsmay be administered by different routes. For example, the BPI proteinproduct may be administered intravenously while the antibiotics areadministered intramuscularly, intravenously, subcutaneously, orally orintraperitoneally. Alternatively, the BPI protein product may beadministered intraperitoneally while the antibiotics are administeredintraperitoneally or intravenously, or the BPI protein product may beadministered in an aerosolized or nebulized form while the antibioticsare administered, e.g., intravenously. The BPI protein product andantibiotics are preferably both administered intravenously. The BPIprotein product and antibiotics may be given sequentially in the sameintravenous line, after an intermediate flush, or may be given indifferent intravenous lines. The BPI protein product and antibiotics maybe administered simultaneously or sequentially, as long as they aregiven in a manner sufficient to allow both agents to achieve effectiveconcentrations at the site of infection.

Concurrent administration of BPI protein product and antibiotic isexpected to provide more effective treatment of grain-positive bacterialinfection. Concurrent administration of the two agents may providegreater therapeutic effects in vivo than either agent provides whenadministered singly. It may permit a reduction in the dosage of one orboth agents with achievement of a similar therapeutic effect.Alternatively, the concurrent administration may produce a more rapid orcomplete bactericidal/bacteriostatic effect than could be achieved witheither agent alone.

Therapeutic effectiveness is based on a successful clinical outcome, anddoes not require that the antimicrobial agent or agents kill 100% of theorganisms involved in the infection. Success depends on achieving alevel of antibacterial activity at the site of infection that issufficient to inhibit the bacteria in a manner that tips the balance infavor of the host. When host defenses are maximally effective, theantibacterial effect required may be minimal. Reducing organism load byeven one log (a factor of 10) may permit the host's own defenses tocontrol the infection. In addition, augmenting an earlybactericidal/bacteriostatic effect can be more important than long-termbactericidal/bacteriostatic effect. These early events are a significantand critical part of therapeutic success, because they allow time forhost defense mechanisms to activate. Increasing the bactericidal ratemay be particularly important for infections such as meningitis, bone orjoint infections. Stratton, Antibiotics in Laboratory Medicine, 3rd ed.(Lorian, V., Ed.) pp. 849-879, Williams and Wilkins, Baltimore Md.(1991)!.

The effect of BPI protein product to improve the therapeuticeffectiveness of antibiotics in vivo may be demonstrated in in vivoanimal models, or may be predicted on the basis of a variety of in vitrotests, including (1) determinations of the minimum inhibitoryconcentration (MIC) of an antibiotic required to inhibit growth of agram-negative organism for 24 hours, (2) determinations of the effect ofan antibiotic on the kinetic growth curve of a gram-negative organism,and (3) checkerboard assays of the MIC of serial dilutions of antibioticalone or in combination with serial dilutions of BPI protein product.Exemplary models or tests are described in Eliopoulos and Moellering InAntibiotics in Laboratory Medicine, 3rd ed. (Lorian, V., Ed.) pp.432-492, Williams and Wilkins, Baltimore Md. (1991).

Using in vitro determinations of antibiotic MIC at 24 hours, a BPIprotein product may be shown to reduce the MIC of the antibiotic. Withthis result, it is expected that concurrent administration of the BPIprotein product in vivo will increase susceptibility of thegram-negative organism to the antibiotic. A BPI protein product may alsobe shown to reduce the MIC of an antibiotic from the range in which theorganism is considered clinically resistant to a range in which theorganism is considered clinically susceptible. With this result, it isexpected that concurrent administration in vivo of the BPI proteinproduct with the antibiotic will reverse resistance and effectivelyconvert the antibiotic-resistant organism into an antibiotic-susceptibleorganism.

By measuring the effect of antibiotics on the in vitro growth curves ofgram-negative organisms, in the presence or absence of a BPI proteinproduct, the BPI protein product may be shown to enhance the earlyantibacterial effect of antibiotics at 0-24 hours. Enhancement of earlybactericidal/growth inhibitory effects is important in determiningtherapeutic outcome.

BPI protein product is thought to interact with a variety of hostdefense elements present in whole blood or serum, including complement,p15 and LBP, and other cells and components of the immune system. Suchinteractions may result in potentiation of the activities of BPI proteinproduct. Because of these interactions, BPI protein products can beexpected to exert even greater activity in vivo than in vitro. Thus,while in vitro tests are predictive of in vivo utility, absence ofactivity in vitro does not necessarily indicate absence of activity invivo. For example, BPI has been observed to display a greaterbactericidal effect on gram-negative bacteria in whole blood or plasmaassays than in assays using conventional media. Weiss et al., J. Clin.Invest. 90:1122-1130 (1992)!. This may be because conventional in vitrosystems lack the blood elements that facilitate or potentiate BPI'sfunction in vivo, or because conventional media contain higher thanphysiological concentrations of Mueller-Hinton has ˜physiologicalconclusion as serum magnesium and calcium, which are typicallyinhibitors of the antibacterial activity of BPI protein products.Furthermore, in the host, BPI protein product is available to neutralizetranslocation of gram-negative bacteria and concomitant release ofendotoxin, a further clinical benefit not seen in or predicted by invitro tests.

It is also contemplated that the BPI protein product be administeredwith other products that potentiate the bactericidal activity of BPIprotein products. For example, serum complement potentiates thegram-negative bactericidal activity of BPI protein products; thecombination of BPI protein product and serum complement providessynergistic bactericidal/growth inhibitory effects. See, e.g., Ooi etal. J. Biol. Chem., 265: 15956 (1990) and Levy et al. J. Biol. Chem.,268: 6038-6083 (1993) which address naturally-occurring 15 kD proteinspotentiating BPI antibacterial activity. See also co-owned, co-pendingPCT Application No. US94/07834 filed Jul. 13, 1994, which corresponds toU.S. patent application Ser. No. 08/274,303 filed Jul. 11, 1994 as acontinuation-in-part of U.S. patent application Ser. No. 08/093,201filed Jul. 14, 1993. These applications, which are all incorporatedherein by reference, describe methods for potentiating gram-negativebactericidal activity of BPI protein products by administeringlipopolysaccharide binding protein (LBP) and LBP protein products. LBPprotein derivatives and derivative hybrids which lack CD-14immunostimulatory properties are described in.

BPI is commonly thought to be non-cytotoxic for many types ofgram-positive bacteria. This lack of toxicity may be due primarily tothe low affinity of BPI for the gram-positive cell wall and the abilityof the cell wall to protect the cytoplasmic membrane from prolongedexposure to and subsequent damage by BPI. Because prokaryoticcytoplasmic membranes from both gram-positive and gram-negative bacteriamay have similar structures that are distinct from those of eukaryoticmembranes, BPI protein products may also be cytotoxic for gram-positivebacteria if the cell wall is removed or damaged or if the BPI proteinproducts can be targeted to the cell surface with high affinity.

Without being bound by a theory of the invention, it is believed thatBPI protein product may have several mechanisms of action. BPI proteinproduct may act directly on the cell walls of gram-positive bacteria bybinding to LPS-like molecules such as cell wall peptidoglycans andteichoic acid. If BPI is allowed to reach the inner cytoplasmicmembrane, the amphipathic nature of BPI may allow it to penetrate thecytoplasmic membrane and exert a bactericidal effect. Thus, agents thatact on or disrupt the cell walls of bacteria such as antibiotics,detergents or surfactants, anti-peptidoglycan antibodies,anti-lipoteichoic acid antibodies and lysozyme, may potentiate theactivity of BPI by allowing access to the inner cytoplasmic membrane.

Likewise, BPI's action on the inner cytoplasmic membrane of bacteria maypotentiate the action of antibiotics by allowing penetration of theantibiotic through the inner membrane, thus permitting it to affect thebacterial biochemical machinery. Moreover, because gram-positivebacterial infection may cause stress-induced translocation of bowelflora and/or LPS, BPI may also act beneficially by killing thegram-negative bacteria and neutralizing the LPS.

BPI protein product, through its heparin-binding ability, may interferewith the binding of gram-positive bacteria to the extracellular matrix(ECM) and to host cells. It has been previously shown that a number oforganisms, including the gram-positive bacteria Staphylococcus aureus,Streptococcus mutans and Streptococcus pyogenes (Group A strep), expressheparin-binding receptors. These heparin-binding receptors are believedto mediate binding of organisms to heparin-like molecules in the ECM andon host cells, e.g., endothelial cells. Heparin may also act as a bridgethat mediates adhesion of organisms to host cells that have heparinreceptors.

Finally, BPI protein product may bind to gram-positive bacterial cellwall components, such as peptidoglycans or teichoic acid, and therebyneutralize the action of these cell wall components in inducinggram-positive sepsis. These cell wall components are believed to play arole in gram-positive sepsis and septic shock. Both peptidoglycans andteichoic acids can activate the alternate complement pathway.Gram-positive bacterial cell wall components can also elicit productionof cytokines involved in sepsis, including TNF, IL-1 and IL-6. Bone, J.Critical Care, 8:51-59 (1993); Bone, Arch. Intern. Med., 154:26-34(1994).! Highly purified gram-positive bacterial cell wall preparations(in which the covalent linkages and structure of peptidoglycan andteichoic acid chains remain unaltered) have been shown to stimulate theproduction of TNF-α and IL-6 by human monocytes. Heumann et al., Infect.Immun., 62:2715-2721 (1994).!

An advantage provided by the present invention is the ability to providemore effective treatment of gram-positive bacterial infections byimproving the therapeutic effectiveness of antibiotics againstgram-positive organisms. This allows use of lower concentrations ofhighly toxic or very expensive antibiotics, such as the aminoglycosides,vancomycin, rifampin, lincomycin, chloramphenicol, and thefluoroquinolones. Because the use of some antibiotics is limited bytheir systemic toxicity or prohibitive cost, lowering the concentrationof antibiotic required for therapeutic effectiveness reduces toxicityand/or cost of treatment, and thus allows wider use of the antibiotic.Another advantage is the ability to treat gram-positive organisms thatare normally resistant to one or more antibiotics. The present inventionmay also provide quality of life benefits due to, e.g., decreasedduration of therapy, reduced stay in intensive care units or reducedstay overall in the hospital, with the concomitant reduced risk ofserious nosocomial (hospital-acquired) infections.

The invention also provides improved methods of in vitro decontaminationof fluids and surfaces contaminated with gram-positive bacteria bycontacting the bacteria with BPI protein product alone, or incombination with one or more antibiotics. The amounts of BPI proteinproduct and antibiotics used are amounts that are separately sufficientfor bactericidal/growth inhibitory effects, or amounts sufficient tohave additive or synergistic bactericidal/growth inhibitory effects.These methods can be used in a variety of in vitro applicationsincluding sterilization of surgical and other medical equipment andimplantable devices, including prosthetic joints. These methods can alsobe used for in situ sterilization of indwelling invasive devices such asintravenous lines and catheters, which are often foci of infection.

The invention further provides pharmaceutical compositions for treatmentof gram-positive bacterial infection comprising BPI protein product incombination with an antibiotic lacking gram-negative bactericidalactivity, such as lincomycin and vancomycin. The pharmaceuticalcomposition can optionally comprise a pharmaceutically acceptablediluent, adjuvant or carrier. As another aspect of the invention,antiseptic bactericidal compositions are provided which comprise a BPIprotein product alone, or in combination with an antibiotic.

"LBP protein derivatives" includes natural, synthetic and recombinantlyproduced polypeptides comprising a portion of the amino acid sequence ofLipopolysaccharide Binding Protein (LBP) holoprotein, and which arecharacterized by the ability to bind to LPS but which lack the carboxyterminal-associated immunostimulatory element(s) characteristic of theLBP holoprotein and thus lack the CD14-mediated immunostimulatoryactivity characteristic of LBP holoprotein. LBP derivative proteins aredescribed in detail in PCT Application No. US94/06931 filed Jun. 17,1994, which corresponds to co-owned, co-pending U.S. patent applicationSer. No. 08/261,660, filed Jun. 17, 1994 as a continuation-in-part ofU.S. patent application Ser. No. 08/079,510, filed Jun. 17, 1993, thedisclosures of all of which are incorporated herein by reference.Preferred LBP protein derivatives include N-terminal LBP fragmentshaving a molecular weight of about 25 kD or less. One LBP N-terminalfragment is characterized by the amino acid sequence of the first 197amino acids of the amino-terminus of LBP set out in SEQ ID NOS: 97 and98, and is designated LBP₂₅. LBP protein derivatives also includepolypeptides comprising part or all of one or more of three regions(defined by LBP amino acid sequences 17-45, 65-99 and 141-167)corresponding (by reason of amino acid holology) to LPS binding regionsof BPI (comprising amino acid sequences 17-45, 65-99 and 142-169). LBPprotein derivatives also include polypeptides comprising a portion ofthe amino acid sequence of LBP holoprotein, wherein amino acids areadded, deleted, or replaced, and wherein the LBP protein derivativemaintains LPS-binding activity but lacks CD14-mediated immunostimulatoryactivity. One such LBP derivative is that wherein the alanine residue atposition 131 of the LBP₂₅ polypeptide fragment is substituted with acysteine residue.

LBP derivatives include LBP derivative hybrid proteins comprising aportion of the amino acid sequence of LBP and a portion of at least oneother polypeptide, such as BPI protein or immunoglobulin chain. One suchderivative is an LBP/BPI hybrid protein (LBP(1-197)/BPI(200-456)), whichcomprises amino acid residues 1-197 of LBP followed in sequence by aminoacid residues 1-199 of BPI. Other LBP hybrid proteins comprise LBP aminoacid sequences into which all or portions of LPS binding domains ofother LPS binding proteins (such as BPI) have been inserted orsubstituted.

As used herein, "BPI protein product" includes naturally andrecombinantly produced BPI protein; natural, synthetic, and recombinantbiologically active polypeptide fragments of BPI protein; biologicallyactive polypeptide variants of BPI protein or fragments thereof,including hybrid fusion proteins and dimers; biologically activepolypeptide analogs of BPI protein or fragments or variants thereof,including cysteine-substituted analogs; and BPI-derived peptides. TheBPI protein products administered according to this invention may begenerated and/or isolated by any means known in the art. U.S. Pat. No.5,198,541, the disclosure of which is incorporated herein by reference,discloses recombinant genes encoding and methods for expression of BPIproteins including recombinant BPI holoprotein, referred to as rBPI₅₀and recombinant fragments of BPI. Co-owned, copending U.S. patentapplication Ser. No. 07/885,501 and a continuation-in-part thereof, U.S.patent application Ser. No. 08/072,063 filed May 19, 1993 andcorresponding PCT Application No. 93/04752 filed May 19, 1993, which areall incorporated herein by reference, disclose novel methods for thepurification of recombinant BPI protein products expressed in andsecreted from genetically transformed mammalian host cells in cultureand discloses how one may produce large quantities of recombinant BPIproducts suitable for incorporation into stable, homogeneouspharmaceutical preparations.

Biologically active fragments of BPI (BPI fragments) includebiologically active molecules that have the same or similar amino acidsequence as a natural human BPI holoprotein, except that the fragmentmolecule lacks amino-terminal amino acids, internal amino acids, and/orcarboxy-terminal amino acids of the holoprotein. Nonlimiting examples ofsuch fragments include a N-terminal fragment of natural human BPI ofapproximately 25 kD, described in Ooi et al., J. Exp. Med., 174:649(1991), and the recombinant expression product of DNA encodingN-terminal amino acids from 1 to about 193 or 199 of natural human BPI,described in Gazzano-Santoro et al., Infect. Immun. 60:4754-4761 (1992),and referred to as rBPI₂₃. In that publication, an expression vector wasused as a source of DNA encoding a recombinant expression product(rBPI₂₃) having the 31-residue signal sequence and the first 199 aminoacids of the N-terminus of the mature human BPI, as set out in FIG. 1 ofGray et al., supra, except that valine at position 151 is specified byGTG rather than GTC and residue 185 is glutamic acid (specified by GAG)rather than lysine (specified by AAG). Recombinant holoprotein (rBPI)has also been produced having the sequence (SEQ ID NOS: 145 and 146) setout in FIG. 1 of Gray et al., supra, with the exceptions noted forrBPI₂₃ and with the exception that residue 417 is alanine (specified byGCT) rather than valine (specified by GTT). Other examples includedimeric forms of BPI fragments, as described in co-owned and co-pendingU.S. patent application Ser. No. 08/212,132, filed Mar. 11, 1994, nowU.S. Pat. No. 5,447,913, and corresponding PCT Application No.PCT/US95/03125, the disclosures of which are incorporated herein byreference. Preferred dimeric products include dimeric BPI proteinproducts wherein the monomers are amino-terminal BPI fragments havingthe N-terminal residues from about 1 to 175 to about 1 to 199 of BPIholoprotein. A particularly preferred dimeric product is the dimericform of the BPI fragment having N-terminal residues 1 through 193,designated rBPI₄₂ dimer.

Biologically active variants of BPI (BPI variants) include but are notlimited to recombinant hybrid fusion proteins, comprising BPIholoprotein or biologically active fragment thereof and at least aportion of at least one other polypeptide, and dimeric forms of BPIvariants. Examples of such hybrid fusion proteins and dimeric forms aredescribed by Theofan et al. in co-owned, copending U.S. patentapplication Ser. No. 07/885,911, and a continuation-in-part applicationthereof, U.S. patent application Ser. No. 08/064,693 filed May 19, 1993and corresponding PCT Application No. US93/04754 filed May 19, 1993,which are all incorporated herein by reference and include hybrid fusionproteins comprising, at the amino-terminal end, a BPI protein or abiologically active fragment thereof and, at the carboxy-terminal end,at least one constant domain of an immunoglobulin heavy chain or allelicvariant thereof. Another example of such a hybrid fusion protein is therecombinant expression product of DNA encoding amino acids 1 through 199of BPI joined to DNA encoding amino acids 198 through 456 of LBP,designated BPI(1-199)-LBP(198-456) hybrid, is described in PCTApplication No. US94/06931 filed Jun. 17, 1994, which corresponds toco-owned, co-pending U.S. patent application Ser. No. 08/261,660, filedJun. 17, 1994 as a continuation-in-part of U.S. patent application Ser.No. 08/079,510, filed Jun. 17, 1993, the disclosures of all of which areincorporated herein by reference.

Biologically active analogs of BPI (BPI analogs) include but are notlimited to BPI protein products wherein one or more amino acid residueshave been replaced by a different amino acid. For example, co-owned,copending U.S. patent application Ser. No. 08/013,801 filed Feb. 2, 1993and corresponding PCT Application No. US94/01235 filed Feb. 2, 1994, thedisclosures of which are incorporated herein by reference, disclosespolypeptide analogs of BPI and BPI fragments wherein a cysteine residueis replaced by a different amino acid. A preferred BPI protein productdescribed by this application is the expression product of DNA encodingfrom amino acid 1 to approximately 193 or 199 of the N-terminal aminoacids of BPI holoprotein, but wherein the cysteine at residue number 132is substituted with alanine and is designated rBPI₂₁ Δcys or rBPI₂₁.Other examples include dimeric forms of BPI analogs; e.g. co-owned andco-pending U.S. patent application Ser. No. 08/212,132 filed Mar. 11,1994, now U.S. Pat. No. 5,447,913 and corresponding PCT Application No.PCT/US95/03125, the disclosures of which are incorporated herein byreference.

Other BPI protein products useful according to the methods of theinvention are peptides derived from or based on BPI produced byrecombinant or synthetic means (BPI-derived peptides), such as thosedescribed in co-owned and copending PCT Application No. US94/10427 filedSep. 15, 1994, which corresponds to U.S. patent application Ser. No.08/306,473 filed Sep. 15, 1994, and PCT Application No. US94/02465 filedMar. 11, 1994, which corresponds to U.S. patent application Ser. No.08/209,762, filed Mar. 11, 1994, which is a continuation-in-part of U.S.patent application Ser. No. 08/183,222, filed Jan. 14, 1994, which is acontinuation-in-part of U.S. patent application Ser. No. 08/093,202filed Jul. 15, 1993 (for which the corresponding internationalapplication is PCT Application No. US94/02401 filed Mar. 11, 1994),which is a continuation-in-part of U.S. patent application Ser. No.08/030,644 filed Mar. 12, 1993, the disclosures of all of which areincorporated herein by reference.

Presently preferred BPI protein products include recombinantly-producedN-terminal fragments of BPI, especially those having a molecular weightof approximately between 21 to 25 kD such as rBPI₂₃ or rBPI₂₁, ordimeric forms of these N-terminal fragments (e.g., rBPI₄₂ dimer).Additionally, preferred BPI protein products include rBPI₅₀ andBPI-derived peptides.

The administration of BPI protein products is preferably accomplishedwith a pharmaceutical composition comprising a BPI protein product and apharmaceutically acceptable diluent, adjuvant, or carrier. The BPIprotein product may be administered without or in conjunction with knownsurfactants, other chemotherapeutic agents or additional knownanti-microbial agents. A preferred pharmaceutical composition containingBPI protein products (e.g., rBPI₅₀, rBPI₂₃) comprises the BPI proteinproduct at a concentration of 1 mg/ml in citrate buffered saline (5 or20 mM citrate, 150 mM NaCl, pH 5.0) comprising 0.1% by weight ofpoloxamer 188 (Pluronic F-68, BASF Wyandotte, Parsippany, N.J.) and0.002% by weight of polysorbate 80 (Tween 80, ICI Americas Inc.,Wilmington, Del.). Another preferred pharmaceutical compositioncontaining BPI protein products (e.g., rBPI₂₁) comprises the BPI proteinproduct at a concentration of 2 mg/ml in 5 mM citrate, 150 mM NaCl, 0.2%poloxamer 188 and 0.002% polysorbate 80. Such preferred combinations aredescribed in co-owned, co-pending PCT Application No. US94/01239 filedFeb. 2, 1994, which corresponds to U.S. patent application Ser. No.08/190,869 filed Feb. 2, 1994 and U.S. patent application Ser. No.08/012,360 filed Feb. 2, 1993, the disclosures of all of which areincorporated herein by reference.

Suitable antibiotics, and therapeutically effective concentrationsthereof when administered with BPI protein products, may be determinedin in vivo models or according to in vitro tests, for example, the invitro minimum inhibitory concentration (MIC) and in vivo mouseperitonitis or rabbit bacteremia assays taught herein. Suitableantibiotics are antibiotics that act on the bacterial cell wall, cellmembrane, protein metabolism or nucleic acid metabolism. These wouldinclude antibiotics or combinations of antibiotics from the followingclasses: β-lactam antibiotics with or without β-lactamase inhibitors,aminoglycosides, tetracyclines, sulfonamides and trimethoprim,vancomycin, macrolides, fluoroquinolones and quinolones, polymyxins, andother antibiotics. Dosage and administration of suitable antibiotics areknown in the art, and briefly summarized below.

Penicillins

When a BPI protein product is concurrently administered with apenicillin, for treatment of a gram-negative bacterial infection, theBPI protein product is generally given parenterally in doses rangingfrom 1 μg/kg to 100 mg/kg daily, and preferably at doses ranging from 1mg/kg to 20 mg/kg daily. The penicillin is generally given in dosesranging from 1 μg/kg to 750 mg/kg daily, preferably not to exceed 24grams daily for adults (or 600 mg/kg daily for children), and ispreferably administered as follows:

Penicillin G is preferably administered parenterally to adults in dosesranging from 600,000 to 1,000,000 units per day. In conventionaladministration, it is effective largely against gram-positive organisms.For treatment of pneumococcal meningitis, penicillin G is administeredin doses of 20-24 million units daily, in divided doses every 2 or 3hours. For children, the preferred parenteral dose of penicillin G is300,000 to 1,000,000 units per day. One unit of penicillin G contains0.6 μg of pure sodium penicillin G (i.e., 1 mg is 1667 units).

Amoxicillin may be administered parenterally to adults in doses rangingfrom 750 mg to 1.5 grams per day, in 3 equally divided doses. Forchildren, preferred parenteral doses of amoxicillin range from 20 to 40mg/kg per day in 3 equally divided doses. Amoxicillin is also availablein combination with clavulanic acid, a β-lactamase inhibitor. A 250 mgdose of the combination drug amoxicillin/clavulanate will contain 250 mgof amoxicillin and either 125 or 62.5 mg of clavulanic acid. Thecombination is preferably administered to adults orally in doses of 750mg per day divided into 3 equal doses every 8 hours, with a preferreddose of 1.5 grams per day for severe infections, given in 3 equallydivided doses. In children, the preferred oral dose is 20 to 40 mg/kgper day in 3 equally divided doses.

Ampicillin is preferably administered parenterally to adults in doses of6 to 12 grams per day for severe infections, in 3 to 4 equally divideddoses. In children, the preferred parenteral dose of ampicillin is 50 to200 mg/kg per day in 3 to 4 equally divided doses. Larger doses of up to400 mg/kg per day, for children, or 12 grams per day, for adults, may beadministered parenterally for treatment of meningitis. Ampicillin isalso available in combination with sulbactam, a β-lactamase inhibitor.Each 1.5 gram dose of ampicillin/sulbactam contains 1 gram of ampicillinand 0.5 grams of sulbactam. The combination is preferably administeredparenterally to adults in doses of 6 to 12 grams per day divided into 4equal doses every 6 hours, not to exceed a total of 12 grams per day.

Azlocillin is preferably administered parenterally to adults in doses of8 to 18 grams per day, given in 4 to 6 equally divided doses.

Carbenicillin is preferably administered parenterally to adults in dosesof 30 to 40 grams per day, given by continuous infusion or in 4 to 6equally divided doses. Daily doses of up to 600 mg/kg have been used totreat children with life-threatening infections.

Mezlocillin is preferably administered to adults parenterally in dosesof 100 to 300 mg/kg per day, given in 4 to 6 equally divided doses. Theusual dose is 16 to 18 grams per day; for life threatening infections,350 mg/kg per day may be administered, but in doses not to exceed 24grams per day given in 6 equally divided doses every 4 hours. Forchildren, the preferred parenteral dose of mezlocillin is 150 to 300mg/kg per day.

Nafcillin is preferably administered intravenously to adults in doses of3 grams per day, given in 6 equally divided doses every 4 hours, withdoubled doses for very severe infections. In conventionaladministration, it is effective largely against gram-positive organisms.In children, the preferred parenteral dose is 20 to 50 mg/kg per day, in2 equally divided doses every 12 hours. The preferred oral dose fornafcillin ranges from 1 gram per day to 6 grams per day in 4 to 6divided doses.

Oxacillin is preferably administered parenterally to adults in doses of2 to 12 grams per day, in 4 to 6 equally divided doses. In conventionaladministration, it is effective largely against gram-positive organisms.In children, oxacillin is preferably administered in doses of 100 to 300mg/kg per day.

Piperacillin is preferably administered parenterally to adults in dosesranging from 100 mg/kg, or 6 grams per day, in 2 to 4 equally divideddoses, up to a maximum of 24 grams per day, in 4 to 6 equally divideddoses. Higher doses have been used without serious adverse effects.

Ticarcillin is preferably administered parenterally to adults in dosesranging from 4 grams per day to 18 grams per day administered in 4 to 6equally divided doses. The usual dose is 200 to 300 mg/kg per day. Forchildren, the preferred parenteral dose of ticarcillin ranges from 50mg/kg per day to 300 mg/kg per day, given in 3, 4 or 6 equally divideddoses. The combination ticarcillin/clavulanate is preferablyadministered parenterally to adults in doses of 200 to 300 mg/kg per day(based on ticarcillin content), in 4 to 6 equally divided doses. Foradults, the usual dose is 3.1 grams (which contains 3 grams ofticarcillin and 100 mg of clavulanic acid) every 4 to 6 hours. Thecombination is also available in a dose of 3.2 grams, which contains 3grams of ticarcillin and 200 mg of clavulanic acid.

In general, it is desirable to limit each intramuscular injection of apenicillin or cephalosporin to 2 grams; larger doses should beadministered by multiple injections in different large muscle masses.

Cephalosporins

When a BPI protein product is concurrently administered with acephalosporin, for treatment of a gram-negative bacterial infection, theBPI protein product is generally given parenterally in doses rangingfrom 1 μg/kg to 100 mg/kg daily, and preferably at doses ranging from 1mg/kg to 20 mg/kg daily. The cephalosporin is generally given in dosesranging from 1 μg/kg to 500 mg/kg daily, preferably not to exceed 16grams daily, and is preferably administered as follows:

Cefamandole is preferably administered parenterally to adults in dosesranging from 1.5 grams per day, given in 3 equally divided doses every 8hours, to 12 grams per day for life-threatening infections, given in 6equally divided doses every 4 hours. In children, cefamandole ispreferably administered in doses ranging from 50 to 150 mg/kg per day,in 3 to 6 equally divided doses, not to exceed a total of 12 grams perday.

Cefazolin is preferably administered parenterally to adults in doses of750 mg per day, given in 3 equally divided doses every 8 hours. Insevere, life-threatening infections, it may be administered at doses of6 grams per day divided into 4 equal doses every 6 hours; in rareinstances, up to 12 grams per day have been used. In children, thepreferred parenteral dose of cefazolin is 20 to 50 mg/kg per day,divided into 3 or 4 equal doses, with 100 mg/kg per day administered forsevere infections.

Cefonicid is preferably administered parenterally to adults in dosesranging from 500 mg once daily, to 2 grams once daily forlife-threatening infections. For intramuscular administration, a 2 gramdose should be divided into two 1-gram injections.

Cefoperazone is preferably administered parenterally to adults in dosesranging from 2 grams per day, given in 2 equally divided doses every 12hours, to 12 grams per day for severe infections, given in 2, 3 or 4equally divided doses. Doses up to 16 grams per day have beenadministered without complications.

Cefotetan is preferably administered parenterally to adults in doses of1 to 4 grams per day, in 2 equally divided doses every 12 hours.Cefotetan may be administered in higher doses for life-threateninginfections, not to exceed a total dose of 6 grams per day.

Cefotaxime is preferably administered parenterally to adults in dosesranging from 1 to 12 grams per day, not to exceed 12 grams per day (2grams every 4 hours) for life-threatening infections. In children, theparenteral dose of cefotaxime is preferably 50 to 180 mg/kg, dividedinto 4 to 6 equal doses.

Cefoxitin is preferably administered parenterally to adults in dosesranging from 3 to 12 grams per day, given in 3, 4, or 6 equally divideddoses. In children, cefoxitin is preferably administered parenterally indoses of 80 to 160 mg/kg per day, given in 4 or 6 equally divided doses,not to exceed a total dose of 12 grams per day.

Ceftazidime is preferably administered parenterally to adults in dosesranging from 500 mg per day, given in 2 to 3 equally divided doses(every 8 or 12 hours), up to a maximum of 6 grams per day. In children,ceftazidime is preferably administered intravenously in doses of 30 to50 mg/kg, to a maximum of 6 grams per day.

Ceftizoxime is preferably administered parenterally to adults in dosesranging from 1 gram per day, given in 2 equally divided doses every 12hours, to 12 grams per day for life-threatening infections, given in 3equally divided doses every 8 hours. The usual adult dose is 1 to 2grams every 8 or 12 hours. For children, the preferred parenteral doseis 50 mg/kg every 6 or 8 hours, for a total daily dose of 200 mg/kg.

Ceftriaxone is preferably administered parentally to adults in dosesranging from 1 to 2 grams per day, given in 2 equally divided dosesevery 12 hours. It may be given in higher doses, not to exceed a totalof 4 grams per day. In children, the preferred parenteral dose ofceftriaxone is 50 to 75 mg/kg per day, not to exceed 2 grams per day. Inmeningitis, ceftriaxone may be administered in doses of 100 mg/kg perday, not to exceed 4 grams per day.

Cefuroxime is preferably administered parenterally to adults in dosesranging from 2.25 to 4.5 grams per day, in 3 equally divided doses every8 hours. For life-threatening infections, 6 grams per day may beadministered in 4 equally divided doses every 6 hours, and formeningitis, 9 grams per day may be administered in 3 equally divideddoses every 8 hours. For children, the preferred parenteral dose ofcefuroxime is 50 to 150 mg/kg per day in 3 to 4 equally divided doses,or 240 mg/kg per day for meningitis.

Cephalexin is formulated for oral administration, and is preferablyadministered orally to adults in doses ranging from 1 to 4 grams per dayin 2 to 4 equally divided doses. For children, the preferred dose is 20to 50 mg/kg per day in divided doses, with doses being doubled forsevere infections.

Cephalothin is usually administered parenterally to adults in doses of 8to 12 grams per day.

Other Beta-Lactams

When a BPI protein product is concurrently administered with an imipenemantibiotic, for treatment of a gram-negative bacterial infection, theBPI protein product is generally given parenterally in doses rangingfrom 1 μg/kg to 100 mg/kg daily, and preferably at doses ranging from 1mg/kg to 20 mg/kg daily. The imipenem is generally given in dosesranging from 1 μg/kg to 100 mg/kg daily, and is preferably administeredas follows:

Imipenem is available in combination with cilastatin, an inhibitor ofthe renal dipeptidase enzyme that rapidly inactivates imipenem. Thecombination is preferably administered intramuscularly to adults indoses of 1 to 1.5 grams per day, given in 2 equally divided doses every12 hours. Intramuscular doses exceeding 1.5 grams per day are notrecommended. The combination is preferably administered intravenously indoses ranging from 1 to 4 grams per day, in 4 equally divided dosesevery 6 hours; doses exceeding 50 mg/kg per day, or 4 grams per day, arenot recommended.

When a BPI protein product is concurrently administered with amonobactam antibiotic, for treatment of a gram-negative bacterialinfection, the BPI protein product is generally given parenterally indoses ranging from 1 μg/kg to 100 mg/kg daily, and preferably at dosesranging from 1 mg/kg to 20 mg/kg daily. The monobactam is generallygiven in doses ranging from 1 μg/kg to 200 mg/kg daily, and ispreferably administered as follows:

Aztreonam is preferably administered parenterally to adults in dosesranging from 1 gram per day, given in 2 equally divided doses every 12hours, up to a maximum recommended dose of 8 grams per day in cases oflife-threatening infection, given in 3 or 4 equally divided doses.

Aminoglycosides

When a BPI protein product is concurrently administered with anaminoglycoside, for treatment of a gram-negative bacterial infection,the BPI protein product is generally given parenterally in doses rangingfrom 1 μg/kg to 100 mg/kg daily, and preferably at doses ranging from 1mg/kg to 20 mg/kg daily. The aminoglycoside is generally given in dosesranging from 1 μg/kg to 20 mg/kg daily, preferably not to exceed 15mg/kg daily, and is preferably administered as follows:

When administering aminoglycosides, it is desirable to measure serumpeak and trough concentrations to ensure the adequacy and safety of thedosage. Dosages should generally be adjusted to avoid toxic peak andtrough concentrations. Amikacin is preferably administered parenterallyto adults and children in doses of 15 mg/kg per day, divided into two orthree equal doses every 8 or 12 hours, and not to exceed a total dose of1.5 grams per day. For uncomplicated infections, a dose of 500 mgamikacin per day, in 2 equally divided doses, may be administered.Dosages should be adjusted to avoid prolonged serum peak concentrationsof amikacin above 35 μg/ml and prolonged trough concentrations greaterthan 10 μg/ml.

Gentamicin is preferably administered parenterally to adults in doses of3 mg/kg per day, in three equally divided doses every 8 hours. Forlife-threatening infections, up to 5 mg/kg per day in 3 to 4 equallydivided doses may be administered, but this dosage should be reduced to3 mg/kg per day as soon as clinically indicated. For children,gentamicin is preferably administered parenterally in doses of 6 to 7.5mg/kg per day. Dosages should be adjusted to avoid prolonged serum peakconcentrations of gentamicin above 12 μg/ml and prolonged troughconcentrations greater than 2 μg/ml.

Netilmicin may be administered parenterally to adults in doses rangingfrom 3 mg/kg per day, in 2 equally divided doses every 12 hours, to 6.5mg/kg per day for serious systemic infection, in 2 or 3 equally divideddoses. In children, the preferred parenteral dose is 5.5 to 8 mg/kg perday, in 2 or 3 equally divided doses. Dosages should be adjusted toavoid prolonged serum peak concentrations of netilmicin above 16 μg/mland prolonged serum trough concentrations above 4 μg/ml.

Tobramycin is preferably administered parenterally to adults in doses of3 mg/kg per day, given in three equally divided doses every 8 hours. Forlife-threatening infections, tobramycin may be administered in doses upto 5 mg/kg per day, in 3 or 4 equally divided doses, but this dosageshould be reduced to 3 mg/kg per day as soon as clinically indicated. Inchildren, tobramycin is preferably administered parenterally in doses of6 to 7.5 mg/kg per day. Prolonged serum concentrations of tobramycinabove 12 μg/ml should be avoided, and rising trough levels above 2 μg/mlmay indicate tissue accumulation, which may contribute to toxicity.

Concurrent administration of BPI protein product with theaminoglycosides, including amikacin, gentamicin, netilmicin andtobramycin, may permit a lowering of the dose of these toxic antibioticsnecessary to achieve a therapeutic effect.

Tetracyclines

When a BPI protein product is concurrently administered with atetracycline, for treatment of a gram-negative bacterial infection, theBPI protein product is generally given parenterally in doses rangingfrom 1 μg/kg to 100 mg/kg daily, and preferably at doses ranging from 1mg/kg to 20 mg/kg daily. The tetracycline is generally given in dosesranging from 1 μg/kg to 50 mg/kg daily, and is preferably administeredas follows:

The tetracycline antibiotics are generally administered to adults indoses of 1 to 2 grams per day. An exception is doxycycline, which ispreferably administered intravenously to adults in doses of 100 to 200mg per day, and to children in doses of 2 mg/lb per day. Tetracyclinemay be administered parenterally to adults in doses of 0.5 to 2 gramsper day, in 2 equally divided doses, and to children in doses of 10 to20 mg/kg per day.

Sulfonamides

When a BPI protein product is concurrently administered with asulfonamide or trimethoprim, for treatment of a gram-negative bacterialinfection, the BPI protein product is generally given parenterally indoses ranging from 1 μg/kg to 100 mg/kg daily, and preferably at dosesranging from 1 mg/kg to 20 mg/kg daily. The sulfonamide or trimethoprimis generally given in doses ranging from 1 μg/kg to 150 mg/kg daily,preferably not to exceed a combination dose of 960 mg trimethoprim/4.8 gsulfamethoxazole daily, and is preferably administered as follows:

The combination trimethoprim/sulfamethoxazole is available in aformulation containing a 1:5 ratio of trimethoprim and sulfamethoxazole(e.g., 16 mg trimethoprim and 80 mg sulfamethoxazole). The combinationis preferably administered intravenously to adults or children in dosesof 8 to 10 mg/kg (based on the weight of the trimethoprim component) perday, in 2 to 4 equally divided doses. For Pneumocystis cariniiinfection, the combination can be administered in doses of 20 mg/kg(based on the weight of the trimethoprim component) per day, in 3-4equally divided doses, to a maximum recommended dose of 960 mgtrimethoprim/4.8 g sulfamethoxazole per day. Trimethoprim alone ispreferably administered orally to adults in doses of 200 mg per day.Sulfamethoxazole alone is preferably administered orally to adults indoses of 2 to 3 grams per day, and to children orally in doses of 50 to60 mg/kg per day.

Fluoroquinolones

When a BPI protein product is concurrently administered with afluoroquinolone or quinolone, for treatment of a gram-negative bacterialinfection, the BPI protein product is generally given parenterally indoses ranging from 1 μg/kg to 100 mg/kg daily, and preferably at dosesranging from 1 mg/kg to 20 mg/kg daily. The fluoroquinolone or quinoloneis generally given in doses ranging from 1 μg/kg to 50 mg/kg daily,preferably not to exceed 1 gram daily, and is preferably administered asfollows:

Norfloxacin is preferably administered orally to adults in doses from400 to 800 mg daily, divided into two doses every 12 hours. Cinoxacin ispreferably administered orally to adults in doses of 1 gram per day,given in 2 or 4 equally divided doses. Ciprofloxacin is preferablyadministered to adults intravenously in doses from 400 to 800 mg daily,or orally in doses from 500 to 1500 mg daily, divided into two dosesevery 12 hours. Ofloxacin is preferably administered to adultsintravenously in doses from 400 to 800 mg daily, or orally in doses from400 to 800 mg daily, divided into two doses every 12 hours.

Vancomycin

When a BPI protein product is concurrently administered with vancomycin,for treatment of a gram-negative bacterial infection, the BPI proteinproduct is generally given parenterally in doses ranging from 1 μg/kg to100 mg/kg daily, and preferably at doses ranging from 1 mg/kg to 20mg/kg daily. The vancomycin is generally given in doses ranging from 1mg/kg to 50 mg/kg daily, and is preferably administered parenterally toadults in doses of 2 grams per day, divided into 2 or 4 doses every 6 or12 hours. In children it is preferably administered in doses of 40mg/kg, given in 4 equally divided doses every 6 hours. In conventionaladministration, vancomycin is effective largely against gram-positiveorganisms.

Macrolides

When a BPI protein product is concurrently administered with amacrolide, for treatment of a gram-negative bacterial infection, the BPIprotein product is generally given parenterally in doses ranging from 1μg/kg to 100 mg/kg daily, and preferably at doses ranging from 1 mg/kgto 20 mg/kg daily. The macrolide is generally given in doses rangingfrom 1 μg/kg to 100 mg/kg daily, and is preferably administered asfollows:

Erythromycin is preferably administered intravenously to adults andchildren in doses of 15 to 20 mg/kg per day, given by continuousinfusion or in 4 equally divided doses every 6 hours. Erythromycin canbe administered at doses up to 4 grams per day in cases of very severeinfection.

Clarithromycin is preferably administered orally to adults in doses of500 mg to 1 gram daily, in equally divided doses every 12 hours.

Azithromycin is preferably administered orally to adults at a dose of500 mg on the first day of treatment followed by 250 mg once daily for 4days, for a total dose of 1.5 grams.

Others

When a BPI protein product is concurrently administered with otherantibiotics, for treatment of a gram-negative bacterial infection, theBPI protein product is generally given parenterally in doses rangingfrom 1 μg/kg to 100 mg/kg daily, and preferably at doses ranging from 1mg/kg to 20 mg/kg daily.

Polymyxin B is generally given in doses ranging from 1 unit/kg to 45,000units/kg daily, and is preferably administered intravenously to adultsand children in doses of 15,000 to 25,000 units/kg per day, divided into2 equal doses every 12 hours. It may be administered intramuscularly indoses of 25,000 to 30,000 units/kg per day, although these injectionsare very painful. Doses of polymyxin B as high as 45,000 units/kg perday have been used in limited clinical studies to treat neonates forPseudomonas aeruginosa sepsis. Polymyxin B is the treatment of choicefor P. aeruginosa meningitis, and is preferably administeredintrathecally to adults and older children in doses of 50,000 units oncedaily for 3 to 4 days, followed by 50,000 units every other day; inchildren under two years old, it is administered intrathecally in dosesof 20,000 daily for 3 to 4 days, followed by 25,000 units every otherday.

Chloramphenicol is preferably administered intravenously to adults indoses of 50 mg/kg per day, in 4 equally divided doses; in exceptionalcases, it can be administered in doses up to 100 mg/kg per day. Inchildren, chloramphenicol is preferably administered intravenously indoses of 25 mg/kg per day, although up to 100 mg/kg per day can beadministered in cases of severe infection.

Clindamycin is preferably administered parenterally to adults in dosesranging from 600 mg to 4.8 grams per day, given in 2, 3 or 4 equallydivided doses. It is recommended that the dose in each intramuscularinjection not exceed 600 mg. For children, clindamycin is preferablyadministered parenterally in doses of 15-40 mg/kg per day, given in 3 or4 equally divided doses.

Dosages of all antimicrobial agents should be adjusted in patients withrenal impairment or hepatic insufficiency, due to the reduced metabolismand/or excretion of the drugs in patients with these conditions. Dosesin children should also be reduced, generally according to body weight.Those skilled in the art can readily optimize effective dosages andadministration regimens for the BPI protein product and the antibioticsin concurrent administration.

Other aspects and advantages of the present invention will be understoodupon consideration of the following illustrative examples. Example 1addresses plate assays of the bactericidal effect in vitro of a BPIprotein product on the gram-positive organism Bacillus subtilis. Example2 relates to plate assays of the in vitro growth inhibitory effect of aBPI protein product on the gram-positive organism Staphylococcus aureusand its L-phase variant. Example 3 evaluates the in vitro bactericidaland growth inhibitory effects of a variety of BPI protein products onStaphylococcus aureus, penicillin-treated S. aureus, and the L-phasevariant, as measured in radial diffusion assays, plate assays, proteinsynthesis assays, and broth growth inhibition assays. Examples 4 and 5evaluate the in vitro growth inhibitory effects of a variety of BPIprotein products on L-phase variants of Streptococcus pneumoniae andEnterococcus faecalis, respectively. Example 6 examines the effect of avariety of BPI protein products on the mycoplasma Acholeplasmalaidlawii. Example 7 examines the in vitro growth inhibitory effect ofan LBP-BPI hybrid on a variety of gram-positive L-phase variants.Example 8 relates to radial diffusion assays measuring the growthinhibitory effect of a variety of BPI synthetic peptides on S. aureus.Examples 9-17 address large-scale screening of the bactericidal/growthinhibitory effect and the antibiotic susceptibility-increasing effect ofBPI protein products on a variety of gram-positive organisms: S.pneumoniae (Example 9), Streptococcus pyogenes (Group A strep) (Example10), Streptococcus agalactia (Group B strep) (Example 11), Streptococcusbovis (Example 12), E. faecalis (Example 13), Enterococcus faecium(Example 14), a variety of Enterococcus species (Example 15), S. aureus(Example 16), Staphylococcus epidermidis (Example 17), and variouscoagulase-negative Staphylococcus species (Example 18). Example 19examines the early in vitro bactericidal effect of BPI and selectedantibiotics on Streptococcus pneumoniae, Staphylococcus aureus andEnterococcus faecalis. Example 20 evaluates the effect of a BPI proteinproduct, alone or with antibiotics, on Streptococcus pneumoniae(especially penicillin-resistant strains), Staphylococcus aureus(especially methicillin-resistant strains), Enterococcus (especiallymultiply resistant strains), and Corynebacteria. Example 22 evaluatesthe in vivo effects of BPI protein products alone or in combination withantibiotics in animal models.

EXAMPLE 1 In Vitro Bactericidal Effects of a BPI Protein Product onBacillus subtilis

A plate assay was used to determine the effects of a BPI protein producton the gram-positive organism Bacillus subtilis (ATCC Accession No.6633). The cells were incubated overnight at 37° C. in Brain HeartInfusion (BHI) broth (Difco, Detroit, Mich.), after which the opticaldensity of the cell suspension was adjusted with additional BHI broth toA₆₀₀ =˜1.0 (equivalent to about 5×10⁸ CFU/ml). The cells were washedtwice with 0.9% NaCl and resuspended in D-PBS, pH 7.4, to a cell densityof approximately 1-2×10⁴ CFU/ml. The cells were diluted at a ratio of1:10 in Earl's MEM medium (GIBCO, Grand Island, N.Y.) and 190 μl of thediluted cells (200-400 cells per well) added to low protein-binding 96well plates (Corning, N.Y.). The rBPI₂₃ at varying concentrations wasadded to the wells to achieve final concentrations ranging from 200 to1,000 nM in a total volume of 200 μl in each well, and the plates wereincubated at 37° C. for 30 minutes or 60 minutes. After incubation, 100μl from each well was plated on brain heart infusion agar plates. After24 hours of incubation at 37° C., the number of colonies on each agarplate was counted.

In multiple experiments, rBPI₂₃ displayed clear and reproduciblebactericidal activity at concentrations as low as 200 nM (about 4.5μg/ml). The results of a representative experiment are displayed in FIG.1, which shows that the bactericidal effects of rBPI₂₃ weredose-dependent. The bactericidal effects of rBPI₂₃ were more pronouncedafter 60 minutes of incubation than after 30 minutes.

EXAMPLE 2 In Vitro Effects of a BPI Protein Product on Staphylococcusaureus and Its L-Phase Variant in a Plate Assay

A plate assay was conducted to test the effect of a BPI protein producton the gram-positive organism Staphylococcus aureus (ATCC Accession No.19636) and its L-phase variant (ATCC Accession No. 19640). An L-phasevariant (also called an L-form) lacks a cell wall and must be grown inthe presence of osmotically protective media containing serum (usually 5to 10% horse serum).

S. aureus, or Escherichia coli J5 as a control were grown overnight inheart infusion (HI) broth and tryptone yeast extract (TYE) broth (bothbroths from Difco, Detroit, Mich.), respectively, then diluted 1:100 inthe same broth and grown to mid-log phase (A₆₀₀ =˜0.5). The S. aureusL-phase variant cells were grown on agar plates containing HI brothsupplemented with 3.5% NaCl and 10% horse serum, and colonies wereresuspended in 4 ml of HI broth supplemented with 3.5% NaCl to an A₆₀₀of approximately 0.2 to 0.5. The cell suspensions were spread on agarplates containing HI broth supplemented with 3.5% NaCl and 5% horseserum. rBPI₂₁ was spotted on the surface of the plates in amountsvarying from 0.75 to 3 μg, in 2 μl. No growth inhibition zones wereobserved on the S. aureus plates, but for the L-phase variant, a clearzone of growth inhibition was observed at 3 μg rBPI₂₁, and partial clearzones at 1.5 and 0.75 μg rBPI₂₁. In the control E. coli plate, clearzones were observed at 1.0 and 0.5 μg of rBPI₂₁. No zones of growthinhibition were observed for plates on which buffer alone was spotted.The results indicate that in the absence of a cell wall, growth of thegram-positive bacterium S. aureus is affected by BPI protein products.

EXAMPLE 3 In Vitro Effects of Various BPI Protein Products onStaphylococcus aureus, Penicillin-Treated S. aureus, and S. aureusL-Phase Variant

A. Evaluation in Radial Diffusion Assays

Radial diffusion assays were performed to determine the effect ofvarious BPI protein products on S. aureus (ATCC Accession No. 19636) andpenicillin-treated S. aureus or natural L-phase variants (ATCC AccessionNo. 19640) grown in osmotically protective media. Because penicillininhibits peptidoglycan synthesis, cells that have been contacted withpenicillin lose their cell walls once growth is initiated. If osmoticprotection is provided, a certain percentage of the cells survive andgrow as L-phase variants. This example was directed towards determiningwhether penicillin-treated cells or L-phase variants would besusceptible to BPI protein products because the protective barrier ofthe cell wall was absent.

S. aureus cells were grown overnight to mid-log phase in HI broth andwere adjusted using the same broth to an optical density of A₆₀₀ =˜0.25(equivalent to about 2.5×10⁸ cells/ml) or 2.5 (equivalent to about2.5×10⁹ cells/ml). 80 μl of the cell suspension was added to 8 ml ofmolten agar medium containing 0.8% agarose, HI broth supplemented with3.5% NaCl, and 1,000 units/ml penicillin G (Sigma, St. Louis, Mo.). Theagar was poured into 90 mm plates and allowed to solidify. As a control,the same S. aureus cells were adjusted to an optical density of A₆₀₀=0.25, 0.025, 0.0025 or 0.00025, and added to the same agar medium, butwithout penicillin. Holes, or wells, of 3 mm diameter were prepared inthe 90 mm plates, and 5 μl of two-fold serial dilutions of rBPI₂₁ wereadded to each well. For the penicillin-treated cells, 240, 120, 60, 30,15 and 7.5 picomoles (pmol) per well of rBPI₂₁ were tested, and theplates were incubated for 48 hours at 37° C. in a CO₂ incubator. For thecontrol cells (not grown on penicillin G-containing plates), 400, 200,100 and 50 pmol/well of rBPI₂₁ were tested, and the plates wereincubated for 24 hours. The plates were then examined for zones ofinhibition. The results, shown below in FIG. 2, demonstrate that the BPIprotein product inhibited bacterial growth in a dose-dependent manner;rBPI₂₁ at levels as low as 30 pmol per well was able to inhibit growthof the penicillin-treated S. aureus cells. In contrast, even the highestconcentration of rBPI₂₁ (400 pmol) had no effect on the control S.aureus cells grown without penicillin.

These results show that, while the normal bacterial form of S. aureus isnot susceptible to BPI protein products, it can be rendered susceptibleby penicillin treatment that causes loss of the cell wall due toinhibition of cell wall synthesis. This indicates that gram-positivebacteria are susceptible to BPI protein products if the cell wall isremoved and the cytoplasmic membrane is exposed.

Additional radial diffusion assays were conducted on the natural S.aureus L-phase variant to confirm the above results. The cells weregrown overnight in HI broth supplemented with 3.5% NaCl, 10 mM CaCl₂ and1000 units/ml penicillin G, to an A₆₀₀ =˜0.3. The cells were used eitherundiluted or as a 1:10 dilution. 80 μl of the cell suspension was addedto 8 ml of molten HI agar medium containing HI broth supplemented with3.5% NaCl and 0.8% agarose. The agar was poured into 90 mm plates andallowed to solidify. As a control, a plate using the same medium wasalso prepared using 80 μl of normal S. aureus cells adjusted to A₆₀₀=2.5 with HI broth supplemented with 3.5% NaCl, 10 mM CaCl₂, and 1000μ/ml penicillin G. Holes, or wells, of 3 mm diameter were prepared inthe 90 mm plates, and 5 μl of rBPI₂₁ in two-fold serial dilutions,starting at 5 μg/5 μl (or 240 pmol/5 μl), was added to the wells. Theplates were incubated for 48 hours and then examined for zones of growthinhibition. The results of this experiment, shown in FIG. 3, demonstratethat the L-phase variant is also susceptible to BPI protein products.

Further radial diffusion assays were conducted to evaluate the effect ofvarious BPI protein products, rBPI₂₃, rBPI₂₁, rBPI₅₀, BPI-Immunoglobulinfusion (the expression product of l.c."p"ING4512 described in Theofan,et al., U.S. application Ser. No. 07/885,911 filed May 19, 1992 and Ser.No. 08/064,693 filed May 19, 1993) and rBPI₄₂ dimer (the dimeric form ofN-terminal (1-193) BPI, described in Amons, et al., U.S. applicationSer. No. 08/212,132 filed Mar. 11, 1994) on penicillin-treated S.aureus. S. aureus were grown to log phase in HI broth supplemented with3.5% NaCl, 5 mM CaCl₂ and 1000 units/ml penicillin G, and was adjustedto A₆₀₀ =˜0.025. The cell suspension was added in a 1:100 dilution tomolten HI agar medium containing HI broth supplemented with 3.5% NaCl,0.8% agarose, and 1000 units/ml penicillin G, which was then allowed tosolidify. Serial 2-fold dilutions of each BPI protein product in 5 μlwere added to 3-mm wells prepared in the plates. After 24 hours ofincubation, the plates were examined for zones of growth inhibition. Theresults of these assays, shown in FIG. 4, demonstrate that these BPIprotein products are all effective in inhibiting growth ofpenicillin-treated S. aureus, with BPI-Immunoglobulin fusion and rBPI₄₂dimer being the most potent.

When these experiments were repeated using other species of bacteria,Staphylococcus epidermidis (ATCC Accession No. 35983), Enterococcusfaecalis (ATCC Accession No. 4200), and Streptococcus pneumoniae (ATCCAccession No. 35088), using the BPI protein products rBPI₂₃ (5 μg/well),rBPI₂₁ (10 μg/well), rBPI₄₂ dimer (5 μg/well), rBPI₅₀ (20 μg/well) andrBPI-Ig fusion (5 μg/well), no growth inhibition was observed for any ofthese BPI protein products. These results indicate that in this assay,the normal bacterial forms of these species of bacteria are also notsusceptible to the BPI protein products when used at the concentrationsindicated. Higher concentrations of these BPI protein products mayproduce different susceptibility results.

The effects of additional BPI protein products, BPI-derived peptides, onthe S. aureus bacterial form (ATCC No. 19636) and its correspondingL-phase variant (ATCC No. 19640) were also evaluated in this radialdiffusion assay. The cells were grown to log phase in either HI broth(for the bacterial forms) or HI broth supplemented with 3.5% NaCl, 5 mMCaCl₂ and 1000 U/mL penicillin G (for the L-phase variants). rBPI₄₂dimer and the BPI-derived peptides XMP.13, XMN.30, XMP.48 and XMP.102(as described in Example 8 and Table 1 infra) were dissolved at ˜2 mg/mLin PBS (without calcium and magnesium) and the corrected concentrationdetermined by comparing absorbance at 280 nM with that predicted by theextinction co-efficient. The bacteria were incorporated into agarose, 5μl aliquots of various concentrations of the peptides or rBPI₄₂ dimerwere added to each well, and the plates were incubated at 37° C. for 24hours (bacterial form) or 48 hours (L-phase variants). The results,shown in FIGS. 5 and 6, indicate that XMP.48 was active against thebacterial form and that all of the peptides tested were active againstthe L-phase variants. The rBPI₄₂ dimer appeared to be about ten-foldmore active on a molar basis than any of the peptides, while XMP.13appeared to be the least potent.

The experiment was repeated with BPI-derived peptides XMP.14, XMP.2,XMP.13, XMP.30, XMP.48 and XMP.102 on the L-phase variant andpenicillin-treated bacterial form of S. aureus. The results, shown inFIGS. 7 and 8, establish that XMP.14 was completely inactive against theL-phase variant and XMP.2 was less active than XMP.13 on both theL-phase variant and the penicillin-treated bacterial form.

B. Evaluation in a Plate Assay and a Protein Synthesis Assay with S.aureus L-Phase Variant

Plate assays were performed to confirm the relative potency of rBPI₄₂dimer and examine the time course of its bactericidal effects. Cellswere grown to A₆₀₀ =0.3 (˜3×10⁸ cells/mL) in HI broth supplemented withNaCl to 0.4M and 5 mM CaCl₂, diluted 1:100 into NaCl-supplemented PBS,incubated with rBPI₂₁, rBPI₅₀, rBPI₄₂ dimer or buffer, and plated on HIagar supplemented with NaCl and 1% horse serum. The results showed thatrBPI₄₂ dimer caused up to a 75% reduction in colony forming units (CFUs)relative to buffer-treated cells at 2 hours. Incubation of cells witheither buffer or 50 μg/mL rBPI₄₂ dimer for up to 5 hours before beingplated produced optimal killing by dimer at 3 hours. In these assays,rBPI₄₂ dimer was consistently and significantly more potent than eitherof the monomeric forms, rBPI₅₀ or rBPI₂₁. To further extend thesestudies, the effect of these BPI protein products on protein synthesisof the S. aureus L-phase variant was examined. In these experiments,cells were grown to A₆₀₀ =0.5 (-5×10⁸ cells/mL) in HI broth supplementedwith NaCl to 0.4M and 5 mM CaCl₂, diluted 1:100 into NaCl-supplementedHI broth and incubated with BPI protein product. Following incubation at37° C., 0.4 μL of ¹⁴ C-amino acids was added and incubation wascontinued for 30 min. at 37° C. Incorporation of the ¹⁴ C-amino acidswas linear for at least 30 min. Cells were treated with 3 mL cold 10%TCA to arrest protein synthesis and release free ¹⁴ C-amino acids fromthe cells. The cells were then applied to 0.45 μm-pore size HA Milliporefilters (Millipore, Bedford, Mass.), washed once with 3 mL 10% TCA andthen with 5 mL water. The filters were dried and counted, and thepercentage of ¹⁴ C-amino acids incorporated relative to the control wascalculated. An initial experiment to establish the optimum incubationtime with 50 μg/mL rBPI₄₂ dimer demonstrated that maximal inhibition ofprotein synthesis (˜90%) was achieved by 2 hours. Incubation of cellsfor 2 hours with various concentrations of BPI protein product and thenwith ¹⁴ C-amino acids demonstrated that, as in other assays, rBPI₄₂dimer was significantly more potent than the other BPI protein products.The rBPI₂₁ appeared to be less potent than rBPI₂₃ or rBPI₅₀ atinhibiting protein synthesis. Thus, patterns of growth inhibitionparallel the patterns of inhibition of protein synthesis, with rBPI₄₂dimer being most potent at inhibiting growth and protein synthesis.

C. Evaluation in a Broth Growth Inhibition Assay with S. aureus L-PhaseVariant

Broth growth inhibition assays were performed to determine the effect ofa BPI protein product on the growth of S. aureus L-phase variant cells(ATCC Accession No. 19640). Initial attempts to grow L-phase variantcells in HI broth supplemented with horse serum resulted in poor growthor clumping. The effect of CaCl₂ concentration on the growth of L-phasevariant cells was tested to determine whether the cells could be readilygrown in media supplemented with CaCl₂ instead of horse serum. L-phasevariant cells were inoculated from an overnight culture into 2 ml of HIbroth supplemented with 3.5% NaCl, 1,000 units/ml penicillin G andvarious concentrations of calcium chloride. The cells were incubated at37° C. on a rotary shaker and the optical density (A₆₀₀) was measured atset time points. The results, depicted in FIG. 9, show that 5-10 mMCaCl₂ maintains an optimal rate of cell growth. These results show thatCaCl₂ can be substituted for serum to allow growth of the L-phasevariants in broth.

L-phase variant cells of S. aureus were cultured overnight in HI brothsupplemented with 3.5% NaCl, 10 mm CaCl₂ and 1,000 units/ml penicillinG. The cells were adjusted to an optical density of A₆₀₀ =˜0.025(equivalent to ˜2.5×10⁷ cells/ml) in the same broth and 150 μl of cellswas added to each well of a 96 well plate. Varying concentrations ofrBPI₂₁, diluted in buffer containing 5 mM citrate and 150 mM NaCl to atotal volume of 2.5 μl, were added to the wells. Control wells containedcells with dilution buffer only (no BPI protein product) or with 16μg/ml lipopolysaccharide binding protein (LBP). The cells were incubatedat 37° C. on a rotary shaker and cell growth (A₆₀₀) was followed over 7hours. The results are shown in FIG. 10, which depicts the change inA₆₀₀ over time for varying concentrations of rBPI₂₁, LBP or control withno BPI protein product. The open squares indicate the control, the opendiamonds indicate 20 μg/ml rBPI₂₁, the open circles indicate 10 μg/mlrBPI₂₁, the open triangles indicate 5 μg/ml rBPI₂₁, the diamonds with across inside indicate 1 μg/ml rBPI₂₁ and the open circle with a crossinside indicates 16 μg/ml LBP. The results indicate a dose-dependentinhibitory effect on cell growth by 7 hours, with the decrease inturbidity at 20 μg/ml rBPI₂₁ suggesting that some cell lysis hasoccurred.

Further experiments were performed to confirm this result and to examinethe effect of CaCl₂ on growth inhibition of S. aureus L-phase variantsby BPI protein product. The L-phase variant cells were grown overnightin HI broth supplemented with 3.5% NaCl, 10 mM CaCl₂ and 1,000 units/mlpenicillin G, and were inoculated into 96 well plates to A₆₀₀ =˜0.02 in100 μl of the same broth supplemented with 2.5, 5 or 10 mM CaCl₂. As acontrol, the BPI-susceptible gram-negative organism E. coli J5 wasinoculated from an overnight culture into TYE broth (Difco, Detroit,Mich.). Two-fold serial dilutions of rBPI₂₁, diluted in 5 mm citrate and150 mm NaCl to a total volume of 2.5 μl, were added to each well toyield final rBPI₂₁ concentrations of 50 to 0.77 μg/ml. The cells wereincubated at 37° C. on a rotary shaker and cell growth as measured byA₆₀₀ was followed. The results of these experiments are shown in FIGS.11, 12 and 13 and confirm that BPI protein product inhibits growth ofthe L-phase variant beginning at about 4 hours. The greatest inhibitoryeffect was observed for the cells grown in broth supplemented with 2.5mM CaCl₂. Under these conditions, rBPI₂₁ at concentrations as low as3.12 μg/ml inhibited growth and caused a decrease in absorbance. Higherconcentrations of CaCl₂ (5 and 10 mM) appeared to inhibit the action ofrBPI₂₁ on the cells. The results of the control experiment with E. coliJ5 are shown in FIG. 14. By comparison the E. coli J5 cells wereimmediately inhibited by rBPI₂₁, but only at the three highestconcentrations (50, 25 and 12.5 μg/ml).

EXAMPLE 4 In Vitro Effects of BPI Protein Products on StreptococcusSpecies in a Radial Diffusion Assay

Radial diffusion assays were conducted to evaluate the effect of thesame BPI protein products tested in Example 3, rBPI₂₃, rBPI₂₁, rBPI₅₀,BPI-Immunoglobulin fusion product and rBPI₄₂ dimer, on S. pneumoniae andS. pyogenes. L-phase variants of S. pneumoniae (ATCC Accession No.35088) were initially isolated by plating log-phase bacterial forms onL-phase variant media (HI broth supplemented with 3.5% NaCl, 10% horseserum, and 1000 units/ml penicillin G). The cells grew optimally on agarplates consisting of 1.3% Bacto-agar (Difco, Detroit, Mich.) and BHIbroth supplemented with 2% NaCl and 10% inactivated horse serum. TheL-phase variants were subsequently adapted to grow on this BHI mediumlacking horse serum. Colonies of L-phase variants from the agar plateswere resuspended into BHI broth supplemented with 2% NaCl and adjustedto A₆₀₀ =˜0.025. The cell suspension was added at a 1:100 dilution tomolten BHI agar medium containing 0.8% agarose and BHI brothsupplemented with 2% NaCl and 1000 units/ml penicillin G, which was thenallowed to solidify. Serial 2-fold dilutions of each BPI protein productin unformulated citrate buffer or formulated citrate buffer were addedin 5 μl aliquots to 3-mm wells prepared in the plates. Unformulatedcitrate buffer was 20 mM citrate, pH 5.0, 150 mM NaCl for rBPI₂₃, or 5mM citrate, pH 5.0, 150 mM NaCl for other BPI protein products.Formulated citrate buffer was 20 mM citrate, pH 5.0, 150 mM NaCl, with0.1% poloxamer 188 and 0.002% polysorbate 80 for rBPI₂₃, or 5 mMcitrate, pH 5.0, 150 mM NaCl with 0.2% poloxamer 188 and 0.002%polysorbate 80 for rBPI₂₁, or 5 mM citrate, pH 5.0, 150 mM NaCl with0.1% poloxamer 188 and 0.002% polysorbate 80 for other BPI proteinproducts. The corresponding formulated or unformulated buffer was alsoadded to wells as a control. After 24 hours of incubation at 37° C., theplates were examined for zones of growth inhibition. There were nodifferences in the activity of BPI protien products in formulated vs.unformulated buffer.

The results of one representative assay, shown in FIG. 15, demonstratethat the L-phase variant of S. pneumoniae is susceptible to all of theseBPI protein products, while, the normal bacterial form is not (as shownabove in Example 3). In the figure, a closed square signifies rBPI₂₁, anopen circle indicates rBPI₂₃, an open triangle indicates rBPI₅₀, aclosed inverted triangle indicates unglycosylated rBPI₅₀, an open squareenclosing a plus-sign indicates BPI-Immunoglobulin fusion, and an opensquare signifies rBPI₄₂ dimer. These data show that rBPI₅₀ was the mostpotent inhibitor of growth, followed by rBPI₄₂ dimer andBPI-Immunoglobulin fusion.

Similar results were obtained when the experiment was repeated withpenicillin-treated S. pneumoniae bacterial forms grown in the presenceof osmotic protection. Log-phase cells growing in BHI broth wereconcentrated to A₆₀₀ =˜5.0, added to the same BHI agarose mediumcontaining penicillin as above, tested with serial dilutions of the sameBPI protein products as above, and incubated for 48-72 hours. Thesepenicillin-treated bacteria were also observed to be susceptible to alltested BPI protein products.

The effects of BPI protein products on the bacterial form (ATCC No.25663) and L-phase variant (ATCC No. 27080) of S. pyogenes wereevaluated in this radial diffusion assay. The bacterial form was grownin BHI broth, and the L-phase variant was grown first on ATCC medium No.608 and later in medium 608 supplemented with penicillin G and 10% heatinactivated horse serum. The cells were incorporated into molten BHIagarose (for the bacterial forms), or molten medium 608 agarosesupplemented with penicillin G and 1% heat inactivated horse serum (forthe L-phase variants), and various concentrations of rBPI₂₁, rBPI₂₃,rBPI₅₀ and rBPI₄₂ dimer were tested. As previously observed for theother bacterial species tested, none of the BPI protein products had aninhibitory effect at the concentrations tested on the S. pyogenesbacterial form. The L-phase variant was susceptible to all BPI proteinproducts tested, rBPI₂₁, rBPI₂₃, rBPI₅₀, and rBPI₄₂ dimer, with rBPI₄₂dimer and rBPI₂₃ being most potent and rBPI₅₀ least (this lower potencyof rBPI₅₀ was not seen with other bacterial species).

The effects of additional BPI protein products, BPI-derived peptides, onthe bacterial and L-phase variants of S. pneumoniae (ATCC No. 35088) andS. pyogenes (bacterial form ATCC No. 25663; L-phase variant ATCC No.27080) were also evaluated in this radial diffusion assay. The results,depicted in FIGS. 16 and 17 (for S. pneumoniae L-phase variant andbacterial form, respectively) and FIGS. 18 and 19 (for S. pyogenesL-phase variant and bacterial form, respectively) demonstrate that, ingeneral, these peptides inhibited growth of the L-phase variant in asimilar manner to that observed with the S. aureus L-phase variant.Unlike S. aureus, the S. pyogenes bacterial form was susceptible toXMP.2, XMP.13, XMP.30 and XMP.102 as well as to XMP.48, while the S.pneumoniae bacterial form also appeared to be slightly susceptible tohigh concentrations of XMP.2, XMP.13, XMP.48 and XMP.102, but notXMP.30. However, when the experiment was repeated with XMP.13 andXMP.30, the S. pneumoniae bacterial form appeared to be susceptible toXMP.30. As previously observed, only the L-phase variants were sensitiveto rBPI₄₂ dimer.

EXAMPLE 5 In Vitro Effects of BPI Protein Products on Enterococcusfaecalis L-Phase Variants in a Radial Diffusion Assay

Radial diffusion assays were conducted to evaluate the effect of thesame BPI protein products, used in Example 3, rBPI₂₃, rBPI₂₁, rBPI₅₀,BPI-Immunoglobulin fusion product and rBPI₄₂ dimer, on E. faecalisL-phase variants. L-phase variants of E. faecalis (ATCC Accession No.4200) were initially isolated by plating log-phase bacterial forms onagar medium consisting of 1.3% Bacto-agar (Difco, Detroit, Mich.) andBHI broth supplemented with 0.5% yeast extract, 0.93% NaCl, 9.73%sucrose, 0.025% MgSO₄, 10% heat inactivated horse serum and 1000units/ml penicillin G (Medium 607, ATCC, Rockville, Md.), but also grewwell on BHI agar supplemented only with NaCl, sucrose and horse serum.The cells were subsequently adapted to growth on Medium 607 lackinghorse serum. Colonies of L-phase variants from the agar plates wereresuspended into Medium 607 and adjusted to A₆₀₀ =˜0.025. The cellsuspension was added at a 1:100 dilution to molten BHI agarose mediumcontaining BHI broth, 0.93% NaCl, 9.73% sucrose and 1000 units/mlpenicillin G, which was then allowed to solidify. Serial 2-folddilutions of each BPI protein product in 5 μl were added to 3-mm wellsprepared in the plates. After 24 hours of incubation, the plates wereexamined for zones of growth inhibition. The results of onerepresentative assay, shown in FIG. 20, demonstrate that the L-phasevariant of E. faecalis is susceptible to all of these BPI proteinproducts, while the normal bacterial form is not (as shown above inExample 3). In the figure, a closed square signifies rBPI₂₁, an opencircle indicates rBPI₂₃, an open triangle indicates rBPI₄₂ dimer, anopen square enclosing a plus-sign indicates BPI-Immunoglobulin fusion,and an open diamond signifies rBPI₅₀ Unglycosylated rBPI₅₀ behaved in anidentical manner to rBPI₅₀. These data show that rBPI₄₂ dimer and rBPI₅₀were the most potent inhibitors of growth.

Similar results were obtained when the experiment was repeated withpenicillin-treated E. faecalis bacterial forms grown in the presence ofosmotic protection. Log-phase cells growing in BHI broth wereconcentrated to A₆₀₀ =˜5.0, added to the same BHI agarose medium(consisting of medium 607 with penicillin G but without horse serum asin the radial diffusion assay described above, tested with serialdilutions of rBPI₂₁, rBPI₂₃, BPI-Immunoglobulin fusion and rBPI₄₂ dimer,and incubated for 48-72 hours. These penicillin-treated bacteria werealso observed to be susceptible to all tested BPI protein products. Inthis experiment there were greater differences in activity, with rBPI₄₂dimer and BPI-Immunoglobulin fusion being the most potent inhibitors ofgrowth.

EXAMPLE 6 In Vitro Effects of BPI Protein Products on Mycoplasma inRadial Diffusion Assays

Mycoplasmas are prokaryotes that lack a cell wall. Many members of thisgroup are naturally-occurring, non-pathogenic inhabitants of humans.Mycoplasma pneumoniae, however, is a major cause of primary atypicalpneumonia. The susceptibility of L-phase variants to BPI proteinproducts predicts that mycoplasmas also are susceptible.

Experiments were performed to evaluate the effects of BPI proteinproducts on the mycoplasma Acholeplasma laidlawii, which is relativelyunfastidious and does not require a CO₂ -enriched atmosphere to grow.Cells were grown overnight in HI broth supplemented with 1% PPLO serumfraction (Difco, Detroid, Mich.), incorporated into the same mediumcontaining agarose, and used in radial diffusion assays as describedabove for L-phase variants. BPI protein products (rBPI₂₃, BPI₂₁, BPI₅₀,rBPI₄₂ dimer and purified XMP.30) were added to the wells and the plateswere incubated at 37° C. for 2 days. The results, shown in FIG. 21,demonstrate that all of the BPI forms tested were effective against theAcholeplasma. For rBPI₂₃ and rBPI₂₁, a second zone of reduced growth wasobserved outside of the zone of complete inhibition.

The experiment was repeated with the same BPI protein products andXMP.13, XMP.2 and XMP.14. The results, depicted in FIG. 22, showed thatXMP.13 and XMP.30 formed inhibition zones while XMP.2 and XMP.14 werenot active at the concentrations tested. Additional experiments can beperformed with this Acholeplasma species as well as other mycoplasmaspecies.

EXAMPLE 7 In Vitro Effects of LBP Derivatives on L-Phase Variants of S.aureus, S. pneumoniae and E. faecalis in a Radial Diffusion Assay

BPI protein products and LBP protein derivatives were evaluated in theradial diffusion assays described above for their effect on the growthof L-phase variants of various bacterial species, including S. aureus,S. pneumoniae and E. faecalis. The compounds evaluated were rBPI₅₀,rBPI₂₃, rBPI₂₁, mature LBP protein (LBP₅₀), LBP25,LBP(1-197)/BPI(200-456) hybrid and BPI(1-199)/LBP(198-456) hybrid.

A plasmid encoding the LBP(1-197) BPI(200-456) hybrid was constructed bycombining appropriate portions of the two molecules via a ClaIrestriction site engineered into homologous locations in the DNAencoding the two molecules. The first step necessary for theconstruction of the mammalian expression vector pING4160 was theconstruction of two intermediate plasmids to introduce a ClaIrestriction site by overlap extension PCR mutagenesis at the Ile-Asp atpositions 197-198 in LBP (to generate plasmid pML127) and the Ile-Asp atpositions 199-200 in BPI (to generate plasmid pML126). These were silentmutations which changed the nucleotide sequence only and not the aminoacid sequence. The next step was to combine the amino terminal portionof LBP from pML127 with the carboxyl terminal of BPI from pML126 at thehomologous ClaI sites to generate the intermediate plasmid pML128. Thefinal step was then to subclone the LBP-BPI insert from pML126 into amammalian expression vector to generate pING4160.

To construct plasmid pML127 (LBP with ClaI at 197-198), overlappingprimers were designed to incorporate the changes necessary to encode aClaI recognition site at the desired location. The template was pML125,a plasmid containing an insert encoding full length LBP. The primerswere LBP-10, SEQ. ID. NO: 228, facing downstream, and LBP-11, SEQ. ID.NO: 229, facing upstream. Two separate PCR reactions were carried outwith primer pairs LBP-Bsm, SEQ. ID. NO: 230, facing downstream, andLBP-11, to generate a 600 bp fragment that was then digested with StuIand ClaI to generate a 389 bp fragment, and primer pairs LBP-10 andLBP-8, SEQ. ID. NO: 231, facing upstream, to generate a 328 bp fragmentthat was then digested with ClaI and Bsu36I to generate a 148 bpfragment. The two resulting fragments were then ligated to theBsu36I-StuI vector fragment from pML125 to generate the plasmid pML127.

To construct plasmid pML126 (BPI with ClaI at 199-200), overlappingprimers were designed to incorporate the changes necessary to encode aClaI recognition site at the desired location. The template was pML124,a plasmid containing an insert encoding full length BPI protein. Theprimers were BPI-63, SEQ. ID. NO: 232, facing downstream, and BPI-64,SEQ. ID. NO: 233, facing upstream. Two separate PCR reactions werecarried out with primer pairs BPI-40, SEQ. ID. NO: 234, facingdownstream, and BPI-64, to generate a 260 bp fragment that was thendigested with PmlI and ClaI to generate a 170 bp fragment, and primerpairs BPI-7, SEQ. ID. NO: 235, facing upstream, and BPI-63, to generatea 296 bp fragment that was then digested with ClaI and BstXI to generatea 215 bp fragment. The two resulting fragments were then ligated to theBstXI-PmlI vector fragment from pML124 to generate the plasmid pML126.

To construct pML128, the intermediate plasmid encoding theLBP(1-197)BPI(200-456) hybrid, the 620 bp HindIII-ClaI fragment encodingthe amino terminal region of BPI in the plasmid pML126 was replaced withthe corresponding HindIII-ClaI fragment from pML127 encoding the aminoterminal region of LBP. To construct the mammalian expression vectorpING4160, the 623 bp FspI-Bsu36I fragment of pML128 was ligated to the361 bp SalI-FspI fragment from pING4539 (described in Gazzano-Santaro etal., U.S. application Ser. No. 08/261,660 filed Jun. 17, 1994), whichincludes the LBP signal sequence, and the approximately 8630 bpBsu36I-SalI fragment from pING4321. The latter fragment includessequences encoding part of the carboxyl terminus of BPI and all thevector sequences, which include the CMV promoter and the light chain 3'transcription termination sequences (as described in Ammons et al., U.S.application Ser. No. 08/212,132, filed Mar. 11, 1994).

A plasmid encoding the BPI(1-199) LBP(198-456) hybrid was constructed bycombining appropriate portions of the two molecules via a ClaIrestriction site engineered into homologous locations in the DNAencoding the two molecules. The intermediate plasmid encoding theBPI(1-199)LBP(198-456) hybrid, pML129, was constructed by replacing the620 bp HindIII-ClaI fragment encoding the amino terminal region of LBPin the plasmid pML127 with the corresponding HindIII-ClaI fragment frompML126 encoding the amino terminal region of BPI protein.

To construct the mammalian expression vector pING4161, the 881 bpBstBI-SstII/T4 fragment of pML129, including part of the BPI and all ofthe LBP insert sequence, was ligated to the approximately 8755XhoI/T4-BstBI fragment of pING4147 (described in Gazzano-Santaro et al.,U.S. application Ser. No. 08/261,660 filed Jun. 17, 1994). The latterfragment includes sequences encoding the signal sequence and part of theamino terminus of BPI and all the vector sequences, which include theCMV promoter and the light chain 3' transcription termination sequences.

To obtain the desired hybrid proteins, beads co-cultured with CHO-K1cells transfected with pING4160 or pING4161 as described in U.S. patentappl. Ser. No. 08/072,063 filed May 19, 1993, were washed withapproximately 600 mls of 20 mM sodium acetate, pH 4.0 mM NaCl and then600 mls of the same buffer containing 600 mM NaCl. Protein was eluted intwo steps of 20 mM sodium acetate; the first with 1.0M NaCl and thesecond with 1.5M NaCl, with the majority of the desired protein elutingfrom the S-Sepharose in the 1.0M step. Fractions containing the proteinwere then pooled and diluted to a final NaCl concentration of 300 mMwith the addition of MES buffer, to a final concentration of 20 mM MES,pH 5.0. The diluted material recovered from all cell harvests wascombined, yielding a final volume of approximately 6.5 liters. Thispooled eluate was applied to two columns arranged in a tandem fashion,the first being a 100 ml Q-Sepharose column and the second a 12 mlCM-Spherodex column. The flow through material, which contained thedesired protein, was adjusted to pH 4.0 and loaded in three batches onto a 15 ml S-Sepharose column. Each time the column was washed with 20mM MES, pH 4.0 200 mM NaCl and the bound protein recovered with a stepelution of 20 mM MES, pH 5.5, 1.2M NaCl. The volume of the recoveredprotein was approximately 40 mls. This material was then run on a S-100size exclusion column in 5 mM sodium citrate, pH 5.0, 150 mM NaCl.Column fractions were assayed using Coommassie stained SDS-PAGE andWestern analysis using an anti-LBP primary antibody, and fractionscontaining the desired protein were pooled.

Radial diffusion assays on L-phase variants of S. pneumoniae (ATCCAccession No. 35088) and E. faecalis (ATCC Accession No. 4200) and thenatural L-variant of S. aureus (ATCC Accession No. 19640) were performedas described above in Examples 3, 5 and 6, using varying concentrationsof rBPI₅₀, LBP₅₀, LBP-BPI hybrid, and thaumatin as a control. LBP-BPIhybrid inhibited growth of S. aureus and S. pneumoniae L-phase variants,but not growth of E. faecalis L-phase variant. The LBP-BPI hybrid wasespecially potent against S. pneumoniae, with activity close to that ofrBPI₅₀. Results of one representative assay on S. aureus L-phase variantusing rBPI₂₃, rBPI₅₀, rBPI₂₁, rLBP₂₅, rLBP₅₀, LBP-BPI hybrid and BPI-LBPhybrid are displayed in FIG. 23. The LBP-BPI hybrid was bactericidalwhile rLBP₅₀ and rLBP₂₅ had little or no activity. The BPI-LBP hybridwas also active against the L-phase variant but less so than the LBP-BPIhybrid. As usual, rBPI₄₂ dimer was the most potent molecule against theL-phase variant. These results suggest that the presence of the BPIcarboxyl terminus has enhanced the activity of rLBP₂₅ on prokaryoticmembranes.

Further radial diffusion assays on these three organisms were conductedwith LBP derivatives, LBP peptide 1-2, SEQ. ID NO: 236, and LBP peptide2-1, SEQ. ID NO: 237, and with other BPI protein products, rBPI₄₂ dimer,XMIP.2, XMP.13, XMP.14, XMP.3 and XMP.5. Results of representativeassays on the S. aureus bacterial form and L-phase variant are displayedin FIGS. 24 and 25. At high concentrations, LBP peptide 1-2 had somebactericidal activity against the bacterial form. For the L-phasevariant, LBP peptide 2-1 (amino acids 73-99 from domain II of LBP) wasas potent as XMIP.3 (amino acids 73-99 from domain II of BPI protein).XMP.5, XMP.13 and rBPI₄₂ dimer also displayed bactericidal activityagainst the L-phase variant, with rBPI₄₂ dimer being the most potent ofall molecules tested. Additional assays were performed to evaluate theactivity of LBP peptide 1-2 and LBP peptide 2-1 and other BPI proteinproducts on the E. faecalis bacterial form, the S. pyogenes L-phasevariant of Example 4, and the S. pneumoniae bacterial form and L-phasevariant. LBP peptide 1-2 displayed some bactericidal activity againstthe two bacterial forms at high concentrations (300 pmol/well or more).LBP peptide 1-2 was as potent as rBPI₄₂ dimer against the S. pneumoniaeL-phase variant, and was intermediate in potency between rBPI₄₂ dimerand XMP.13 for the S. pyogenes L-phase variant. LBP peptide 2-1 had nobactericidal activity against the tested bacterial forms or L-phasevariants.

EXAMPLE 8 In Vitro Effects of BPI-Derived Peptides on S. aureus in aRadial Diffusion Assay

This example addresses in vitro screening of BPI protein products,specifically BPI-derived peptides, for antimicrobial activity in aradial diffusion assay. The peptides tested were all prepared accordingto the procedures described in parent U.S. patent application Ser. Nos.08/209,762 and 08/183,222. Briefly summarized, peptides were prepared bysolid phase peptide synthesis according to the methods of Merrifield, J.Am Chem. Soc. 85: 2149 (1963) and Merrifield et al. Anal. Chem., 38:1905-1914 (1966) using an Applied Biosystems, Inc. Model 432 peptidesynthesizer. Peptide design was based in principal part on the discoveryof three functional domains present in the NH₂ -terminal region of theBPI holoprotein domain I comprising BPI amino acids from about position17 to about position 45 (SEQ ID NO: 1); domain II comprising BPI aminoacids from about position 65 to about 99 (SEQ ID NO: 6); and domain IIIcomprising BPI amino acids from about position 142 to about position 169(SEQ ID NO: 12). Peptides included sequences and subsequences of thedomain sequences and variants thereof including linear and branchedchain combination peptides with and without single or multiple aminoacid (including atypical amino acid) substitutions as well as cyclizedpeptides and interdomain sequence peptides. Table 1 below sets outpeptides derived from or based on BPI sequences, as identified by thepeptide number with a prefix XMP or BPI (e.g., XMP.1 or BPI.1, XMP.2 orBPI.2, etc.), SEQ ID NO:, amino acid sequence based on reference toposition within BPI and designation of amino acid substitutions andadditions. Also set out in Table 1 are mass spectroscopy and HPLCestimates of purity of the peptides.

Overnight S. aureus cultures were diluted 1:50 into fresh tryptic soybroth and incubated for 3 hours at 37° C. to attain log phase. Bacteriawere pelleted at 3,000 rpm (1500×g) for 5 minutes in a Beckman J-6Mcentrifuge. 5 ml of 10 mM sodium phosphate buffer, pH 7.4, was added andthe suspension was recentrifuged. Supernatant was decanted and 5 ml offresh buffer was added for an OD₅₇₀ determination. An OD₅₇₀ of 1.0 wasconsidered equivalent to 5×10⁸ CFU/ml. Ten mL of molten underlayeragarose comprising 3% tryptic soy broth, 1% agarose (Pharmacia,Piscataway, N.J.), 0.02% Tween 20, and 10 mM sodium phosphate, at pH 7.4was added to polystyrene tubes and maintained in a 56° C. water bathuntil the addition of bacteria. Tubes were cooled to approximately 45°C., bacteria were added to give a final concentration of 1×10⁶ CFU/ml,and the tubes were mixed again by inverting. The contents were pouredinto level square petri dishes and distributed evenly. The agarosesolidified in less than 30 seconds and had a uniform thickness of about1 mm. A series of wells were punched into the hardened agarose using asterile 3 mm punch attached to a vacuum apparatus.

Peptides to be screened were 2-fold serially diluted in Dulbecco's PBS(D-PBS) starting from a concentration of approximately 1 mg/mL. Five μLof each dilution was added to each well and the plates were incubated at37° C. for 3 hours. An overlayer of 10 mL of molten agarose comprising6% tryptic soy broth, 1% agarose, and 10 mM sodium phosphate, pH 7.4,(at approximately 45° C.) was then added and plates were incubatedovernight at 37° C. Following this overnight incubation, a diluteCoomassie solution was poured into the plates and allowed to stain for24 hours.

Clear zones of growth inhibition around each well were measured withcalipers. The actual area of growth inhibition (mm²) was calculated bysubtracting the area of the well. Table 1 below sets out the results ofthe radial diffusion assays for tested peptides in terms of the numberof μg or picomoles (pmol) of peptide required to establish a 30 mm² areaof growth inhibition.

Peptides XMP.221 through XMP.281 (SEQ ID NOs: 166 through 226) areprepared and tested for anti-fungal activity as described above.

                                      TABLE 1                                     __________________________________________________________________________                                      S. aureus                                                                     μg/                                                                            pmol/                                   Peptide #                MS %                                                                              HPLC %                                                                             30 mm.sup.2                                                                       30 mm.sup.2                             (Seq. ID No.)                                                                         Protein AA Segment                                                                             Purity                                                                            Purity                                                                             zone                                                                              zone                                    __________________________________________________________________________    XMP.1 (4)                                                                             19-33            --  2 Peaks                                                                            N   N                                       XMP.2 (7)                                                                             85-99            64    37.2                                                                             N   N                                       XMP.3 (11)                                                                            73-99            --  17   X   X                                       XMP.4 (3)                                                                             25-46            --  No Peak                                                                            N   N                                       XMP.5 (67)                                                                            142-163          --  18   X   X                                       XMP.7 (54)                                                                            (90-99) × 2                                                                              69  27   >5.00                                                                             >1,891                                  XMP.8 (8)                                                                             90-99            79  Mixtures                                                                           X   X                                       XMP.9 (51)                                                                            95-99, 90-99     --  29   X   X                                       XMP.10 (55, 65)                                                                       94-99, 90-99, 90-99 and                                                                        --  Mixture                                                                            X   X                                               95-99, 90-99, 90-99                                                   XMP.11 (13)                                                                           148-151, 153-161 --  76   X   X                                       XMP.12 (14)                                                                           141-169          --  26   X   X                                       XMP.13 (15)                                                                           148-161          78  69   >5.00                                                                             >2,924                                  XMP.13P (15)                                                                          148-161          100 98   X   X                                       XMP.14 (2)                                                                            21-50            --  --   X   X                                       XMP.15 (16)                                                                           85-99, A @ 85 (I)                                                                              66    57.6                                                                             X   X                                       XMP.16 (17)                                                                           85-99, A @ 86 (K)                                                                              --    84.1                                                                             X   X                                       XMP.17 (18)                                                                           85-99, A @ 87 (I)                                                                              86  77, 67                                                                             X   X                                       XMP.18 (19)                                                                           85-99, A @ 88 (S)                                                                              66  70   X   X                                       XMP.19 (20)                                                                           85-99, A @ 89 (G)                                                                              --  69   X   X                                       XMP.20 (21)                                                                           85-99, A @ 90 (K)                                                                              --  66   X   X                                       XMP.21 (22)                                                                           85-99, A @ 91 (W)                                                                              68    66.8                                                                             X   X                                       XMP.22 (23)                                                                           85-99, A @ 92 (K)                                                                              --  66   X   X                                       XMP.23 (24)                                                                           85-99, A @ 94 (Q)                                                                              --  69   X   X                                       XMP.24 (25)                                                                           85-99, A @ 95 (K)                                                                              --  67   X   X                                       XMP.25 (26)                                                                           85-99, A @ 96 (R)                                                                              --  73   X   X                                       XMP.26 (27)                                                                           85-99, A @ 97 (F)                                                                              --  73   X   X                                       XMP.27 (28)                                                                           85-99, A @ 98 (L)                                                                              --  65   X   X                                       XMP.28 (29)                                                                           85-99, A @ 99 (K)                                                                              --  80   X   X                                       XMP.29 (56)                                                                           (148-161) × 2                                                                            --  26   >5.00                                                                             >1,469                                  XMP.30 (52)                                                                           90-99, 148-161   --  21   3.68                                                                              1,216                                   XMP.30-P (52)                                                                         90-99, 148-161   95  98   X   X                                       XMP.31 (33)                                                                           148-161, A @ 148 (K)                                                                           --  68   X   X                                       XMP.32 (34)                                                                           148-161, A @ 149 (S)                                                                           --  70   X   X                                       XMP.33 (35)                                                                           148-161, A @ 150 (K)                                                                           --  58   X   X                                       XMP.34 (36)                                                                           148-161, A @ 151 (V)                                                                           --  51   X   X                                       XMP.35 (37)                                                                           148-161, A @ 152 (G)                                                                           --  72   X   X                                       XMP.36 (38)                                                                           148-161, A @ 153 (W)                                                                           --  64   X   X                                       XMP.37 (39)                                                                           148-161, A @ 154 (L)                                                                           --  51   X   X                                       XMP.38 (40)                                                                           148-161, A @ 155 (I)                                                                           --  70   X   X                                       XMP.39 (41)                                                                           148-161, A @ 156 (Q)                                                                           --  53   X   X                                       XMP.40 (42)                                                                           148-161, A @ 157 (L)                                                                           --  53   X   X                                       XMP.41 (43)                                                                           148-161, A @ 158 (F)                                                                           --  63   X   X                                       XMP.42 (44)                                                                           148-161, A @ 159 (H)                                                                           --  59   X   X                                       XMP.43 (45)                                                                           148-161, A @ 160 (K)                                                                           --  53   X   X                                       XMP.44 (46)                                                                           148-161, A @ 161 (K)                                                                           --  70   X   X                                       XMP.45 (31)                                                                           85-99, A @ 94 (Q) & 95 (K)                                                                     71  46   >5.00                                                                             >1,697                                  XMP.46 (57)                                                                           (90-99) × 2, A @ 1st 94 (Q) & 95 (K)                                                     67  47   >5.00                                                                             >1,811                                  XMP.47 (58)                                                                           (90-99) × 2, A @ 2d 94 (Q) & 95 (K)                                                      57  34   >5.00                                                                             >2,461                                  XMP.48 (59)                                                                           (90-99) × 2, A @ both 94 (Q) & 95 (K)                                                    68  33   >5.00                                                                             >1,390                                  XMP.54 (5)                                                                            21-35            --  --   X   X                                       XMP.55 (61)                                                                           152-172          --  28   >5.00                                                                             >1,592                                  XMP.56 (47)                                                                           85-99, K @ 94 (Q) &                                                                            --  55   N   N                                               Q @ 95 (K)                                                            XMP.57 (99)                                                                           Cys 85-99 Cys    50  Mixture                                                                            N   N                                       XMP.58 (9)                                                                            Cys-85-99        49    25.7                                                                             N   N                                       XMP.59 (30)                                                                           85-99, A @ 90 (K) & 92 (K)                                                                     56  30   N   N                                       XMP.60 (32)                                                                           85-99, A @ 86 (K) & 99 (K)                                                                     57  78   N   N                                       XMP.61 (48)                                                                           85-99, F @ 91 (W)                                                                              60  60   N   N                                       XMP.63 (53)                                                                           85-99, 148-161   38    31.0                                                                             >5.00                                                                             >1,006                                  XMP.65 Rd (68)                                                                        Cys-85-99-Cys    41  22, 34                                                                             N   N                                       XMP.65 Ox (10)                                                                        Cys-85-99-Cys    --  No Peak                                                                            >5.00                                                                             >3,118                                  XMP.66 (49)                                                                           85-99, W.sub.D @ 91 (W)                                                                        --  70   N   N                                       XMP.67 (50)                                                                           85-99, β-(1-naphthyl)-A                                                                   65  52   N   N                                               @ 91                                                                  XMP.69 (60)                                                                            90-99, A @ 94 (Q) &                                                                           44  54, 40                                                                             3.83                                                                              1,058                                           95 (K)! × 3                                                     XMP.70 (63)                                                                           85-99, β-(3-pyridyl)-A                                                                    66  54   N   N                                               @ 91                                                                  XMP.71 (64)                                                                           A.sub.D -A.sub.D -85-99                                                                        --  60   N   N                                       XMP.72 (66)                                                                           85-99, β-(3-pyridyl)-A                                                                    --  52   N   N                                               @ 97 (F)                                                              XMP.73 (62)                                                                           85-99, F @ 95 (K)                                                                              --  44, 39                                                                             >5.00                                                                             >1,811                                  XMP.74 (70)                                                                           148-161, 90-99   --  29   >5.00                                                                             >2,148                                  XMP.75 (100)                                                                          IKKRAISFLGKKWQK (2-mixed)                                                                      --  32   >5.00                                                                             >2,031                                  XMP.76 (71)                                                                           85-99, F.sub.D @ 95 (K)                                                                        53  39   N   N                                       XMP.77 (72)                                                                           85-99, W @ 95 (K)                                                                              --  38   3.15                                                                              1,684                                   XMP.79 (73)                                                                           85-99, K @ 94 (Q)                                                                              --  48   N   N                                       XMP.80 (74)                                                                           85-99, β-(1-naphthyl)-A                                                                   71  44   4.82                                                                              2,533                                           @ 95 (K)                                                              XMP.81 (75)                                                                           85-99, F @ 94 (Q)                                                                              44  33, 35                                                                             >5.00                                                                             >2,345                                  XMP.82 (76)                                                                           148-161, W @ 158 (F)                                                                           82  58   >5.00                                                                             >1,198                                  XMP.83 (77)                                                                           148-161, β(1-naphthyl)-A                                                                  85  63   >5.00                                                                             >2,034                                          @ 153 (W)                                                             XMP.84 (78)                                                                           85-99, β-(1-naphthyl) A @                                                                 64  50   >5.00                                                                             >2,017                                          91 (W) & F @ 95 (K)                                                   XMP.85 (79)                                                                           148-161, L @ 152 (G)                                                                           79  74   >5.00                                                                             >1,881                                  XMP.86 (80)                                                                           148-161, L @ 156 (Q)                                                                           69  51   >5.00                                                                             >2,048                                  XMP.87 (81)                                                                           148-161, L @ 159 (H)                                                                           79  63   >5.00                                                                             >1,536                                  XMP.88 (82)                                                                           85-99, F @ 94 (Q)                                                                              62  50   >5.00                                                                             >2,380                                          & 95 (K)                                                              XMP.89 (84)                                                                           85-99, β-(1-naphthyl) A @                                                                 66  50   >5.00                                                                             >1,882                                          91 (W) & F @ 94 (Q)                                                   XMP.90 (85)                                                                           85-99, β-(1-naphthyl) A @91 (W),                                                          70  63   >5.00                                                                             >1,863                                          F @ 94 (Q) & 95 (K)                                                   XMP.91 (86)                                                                           148-161, F @ 156 --  31   N   N                                               (Q)                                                                   XMP.92 (87)                                                                           148-161, K @ 156 (Q)                                                                           --  50   N   N                                       XMP.93 (88)                                                                           85-99 148-161 β-(1-naphthyl) A                                                            72  38   >5.00                                                                             >980                                            @ 91 (W), F @ 95 (K)                                                  XMP.94 (89)                                                                           148-161, F @ 159 (H)                                                                           --  59   >5.00                                                                             >922                                    XMP.95 (90)                                                                           148-161, F @ 152 (G)                                                                           --  57   N   N                                       XMP.96 (101)                                                                          148-161, F @ 161 (K)                                                                           --  60   >5.00                                                                             >2,048                                  XMP.97 (92)                                                                           148-161, K @ 152 (G)                                                                           --  67   N   N                                       XMP.98 (83)                                                                           90-99, β-(1-naphthyl) A @ 91 (W),                                                         69  31   N   N                                               F @ 95 (K) + 148-161 F @ 156 (Q)                                      XMP.99 (93)                                                                            90-99, W @ 95   --  --   >5.00                                                                             >1,064                                          (K)! × 3                                                        XMP.100 (94)                                                                          148-161, K @ 152 (G) &                                                                         --  61   N   N                                               156 (Q)                                                               XMP.101 (95)                                                                          (148-161) × 2 K @ 152 (G) & 156 (Q),                                                     --  16   >5.00                                                                             >993                                            F @ 159 (H) & 161 (K)!                                                XMP.102 (96)                                                                          90-99 (F @ 95 (K)) + 148-161 L @                                                               --  16   N   N                                               156 (Q)                                                               XMP.103 (102)                                                                         85-99, W @ 94 (Q)                                                                              --  28   >5.00                                                                             >2,703                                  XMP.104 (103)                                                                         148-161, S @ 156 (Q)                                                                           --  34   >5.00                                                                             >5,569                                  XMP.105 (104)                                                                         85-99, β-(1-naphthyl)-A @ 94 (Q)                                                          58  43   >5.00                                                                             >1,843                                  XMP.106 (105)                                                                         148-161, T @ 156 (Q)                                                                           --  26   N   N                                       XMP.107 (106)                                                                         148-161, W @ 159 (H)                                                                           --  55   N   N                                       XMP.108 (107)                                                                         148-161, W @ 161 (K)                                                                           --  50   >5.00                                                                             >3,219                                  XMP.109 (108)                                                                         148-161, β(1-naphthyl) - A                                                                --  41   N   N                                               @ 158 (F)                                                             XMP.110 (109)                                                                         148-161, β(1-naphthyl) - A                                                                --  56   N   N                                               @ 159 (H)                                                             XMP.111 (110)                                                                         148-161, β(1-naphthyl) - A                                                                --  73   >5.00                                                                             >2,809                                          @ 161 (K)                                                             XMP.112 (111)                                                                         85-99, β(1-naphthyl)A                                                                     --  56   N   N                                               @ 91 (W) & 95 (K)                                                     XMP.113 (112)                                                                         148-161, F @ 157 (L)                                                                           --  46   N   N                                       XMP.114 (113)                                                                         KWQLRSKGKIKIFKA  --  17   N   N                                       XMP.116 (114)                                                                         148-161, K @ 152 (G),                                                                          --  72   N   N                                               β(1-naphthyl)A @ 153 (W)                                         XMP.119 (115)                                                                         85-99, β(1-naphthyl)A @ 91 (W)                                                            --  77   >5.00                                                                             >2,617                                          & 94 (K)                                                              XMP.120 (116)                                                                         85-99, K @ 97 (F)                                                                              --  52   N   N                                       XMP.121 (117)                                                                         85-99, β(1-naphthyl)A @ 94 (Q)                                                            65  35   >5.00                                                                             >2,540                                          & 95 (K)                                                              XMP.122 (118)                                                                         85-99, β(1-naphthyl)A @ 91 (W),                                                           --  46   >5.00                                                                             >2,526                                          94 (Q) & 95 (K)                                                       XMP.123 (119)                                                                         148-161, p-Amino-F @ 156 (Q)                                                                   --  64   N   N                                       XMP.124 (120)                                                                         148-161, K @ 152 (G), W @                                                                      --  67   N   N                                               158 (F)                                                               XMP.125 (121)                                                                         148-161, Y @ 156 (Q)                                                                           --  54   N   N                                       XMP.126 (122)                                                                         148-161, W.sub.D @ 153 (W)                                                                     66  54   N   N                                       XMP.127 (123)                                                                         148-161, F @ 153 (W)                                                                           65  63   N   N                                       XMP.128 (124)                                                                         148-161 F.sub.D @ 153 (W)                                                                      63  51   N   N                                       XMP.129 (125)                                                                         148-161, 1-β(1-naphthyl)A.sub.D @                                                         24  28   N   N                                               153 (W)                                                               XMP.130 (126)                                                                         148-161, 2-β(1-naphthyl)A.sub.D @                                                         55  80   N   N                                               153 (W)                                                               XMP.131 (127)                                                                         148-161, 2-β(1-naphthyl)A.sub.D @                                                         75  60   N   N                                               153 (W)                                                               XMP.132 (128)                                                                         148-161, Pyr-A @ 153 (W)                                                                       49  50   N   N                                       XMP.133 (129)                                                                         148-161, p-Amino-F @ 153 (W)                                                                   63  47   N   N                                       XMP.134 (130)                                                                         148-161, p-Amino-F @ 152 (G)                                                                   --  68   N   N                                       XMP.135 (131)                                                                         148-161, K @ 153 (W)                                                                           --  70   N   N                                       XMP.136 (132)                                                                         85-99, E @ 95 (K)                                                                              --  50   N   N                                       XMP.137 (133)                                                                         Cys-148-161-Cys  --  28   X   X                                       XMP.138 (134)                                                                         148-161, K @ 152 (G), F @ 153 (W)                                                              --  61   N   N                                       XMP.139 (135)                                                                         148-161, Y @ 153 (W)                                                                           --  60   N   N                                       XMP.140 (136)                                                                         90-99 β(1-naphthyl)A @ 94 (Q)                                                             --  26   >5.00                                                                             >1,601                                          & 95 (K) + 104                                                        XMP.141 (137)                                                                         85-99, W @ 97 (F)                                                                              --  50   N   N                                       XMP.142 (138)                                                                         148-161, W @ 157 (L)                                                                           --  57   N   N                                       XMP.143 (139)                                                                         148-161, β(1-naphthyl)A                                                                   --  65   N   N                                               @ 157 (L)                                                             XMP.144 (140)                                                                         148-161, Cyclohexyl-A                                                                          --  60   N   N                                               @ 153 (W)                                                             XMP.145 (141)                                                                         90-99, β(1-naphthyl)A @ 94 (Q)                                                            --  20   X   X                                               & 95 (K) + 148-161                                                    XMP.146 (142)                                                                         148-161, β(1-naphthyl)A @                                                                 --  53   N   N                                               159 (H) & 161 (K)                                                     XMP.147 (143)                                                                         85-99 K @ 96 (R) --  55   N   N                                       XMP.148 (144)                                                                         148-161, β(1-naphthyl)A                                                                   --  62   N   N                                               @ 153 (W) & 159 (H)                                                   XMP.149 (147)                                                                         KWKVFKKIEK + 148-161                                                                           --  27   N   N                                       XMP.150 (148)                                                                         KWAFAKKQKKRLKRQWLKKF                                                                           --  Mixture                                                                            N   N                                       XMP.151 (55)                                                                          94-99, 90-99, 90-99                                                                            --  14   N   N                                       XMP.152 (65)                                                                          95-99, 90-99, 90-99                                                                            --  21   N   N                                       XMP.153 (149)                                                                         (90-99) × 3                                                                              --  17   N   N                                       XMP.154 (150)                                                                         (90-99) × 2, β(1-naphthyl) A                                                        --  31   >5.00                                                                             >1,796                                          1st 94 (Q) & 95 (K)                                                   XMP.155 (151)                                                                         (90-99) × 2, β(1-naphthyl) A                                                        --  23   >5.00                                                                             >1,796                                          @ 2nd 94 (Q) & 94 (K)                                                 XMP.156 (152)                                                                         (90-99) × 2, β(1-naphthyl) A                                                        --  38   N   N                                               both 94 (Q) & 95 (K)                                                  XMP.157 (153)                                                                         (90-99, β(1-naphthyl) A                                                                   --  38   N   N                                               @ 94 (Q) & 95 (K)) × 3                                          0XMP.158 (154)                                                                        85-99, 148-161, β(1-naphthyl) A                                                           --  16   N   N                                               @ 94 (Q) & 95 (K)                                                     XMP.159 (155)                                                                         (90-99, β(1-naphthyl) A                                                                   --  23   >5.00                                                                             >1,590                                          @ 91 (W) & 95 (K)) + 82                                               XMP.160 (156)                                                                         (90-99) × 2, β(1-naphthyl) A @                                                      --  32   >5.00                                                                             >1,782                                          both 91 (W) & 95 (K)                                                  XMP.161 (157)                                                                         148-161, K @ 152 (G) & A @                                                                     --  75   >5.00                                                                             >2,999                                          153 (W)                                                               XMP.162 (158)                                                                         90-99, 148-161, W @ 95 (K)                                                                     --  21   N   N                                       XMP.163 (159)                                                                         (90-99) × 2, W @ both 95 (K)                                                             --  Mixture                                                                            >5.00                                                                             >1,810                                  XMP.164 (160)                                                                         (90-99) × 2, β(1-naphthyl) A @ both                                                 --  46   >5.00                                                                             >1,796                                          94 (Q)                                                                XMP.165 (161)                                                                         (90-99, β(1-naphthyl A @ 91 (W)                                                           --  72   >5.00                                                                             >1,847                                          & F @ 95 (K)) × 2                                               XMP.166 (162)                                                                         148-161, V @ 153 (W)                                                                           --  68   N   N                                       XMP.167 (163)                                                                         90-97            --  56   N   N                                       XMP.168 (164)                                                                         C-90-101-C       --  13   N   N                                       XMP.169 (165)                                                                         C-90-97-C        --  20   >5.00                                                                             >4,974                                  XMP.170 (227)                                                                         90-101           --  69   N   N                                       __________________________________________________________________________     X = Not tested                                                                N = No detectable activity up to 5 μg/well                            

EXAMPLE 9 In Vitro Effects of a Variety of BPI Protein Products Alone orin Combination with Antibiotics on Strains of Streptococcus pneumoniae

This experiment evaluated the in vitro growth inhibitory effects of avariety of BPI protein products, alone or in combination withantibiotics, on clinical isolates of Streptococcus pneumoniae (fromBaxter Microscan® library, Sacramento, Calif.). The BPI proteinproducts' direct growth inhibitory effect and effect on the antibioticsusceptibility of the organisms was determined using Microscan® panelplates (Baxter Diagnostics, Inc., Deerfield, Ill.) that allowsimultaneous determination of minimum inhibitory concentrations for anumber of different antibiotics. Any other antimicrobial panel systemsknown in the art, such as the Pasco (DIFCO, Detroit, Mich.) and Alamar(Alamar, Sacramento, Calif.) systems, may be used instead of theMicroscan® system to assay for activity. Control assays performed withthe Microscan® panel plates confirmed that the formulation buffer forrBPI₂₁ (without rBPI₂₁) had no effect on the antibiotic susceptibilityof a variety of organisms: S. pneumoniae (Microscan library no. 31573),S. pyogenes (Group A) (30403), S. bovis (008-010), S. aureus (052-106)and E. faecalis (011-066).

The antimicrobial susceptibility tests performed on the Microscan® panelplates are miniaturizations of the broth dilution susceptibility test.Antimicrobial agents are serially diluted in Mueller-Hinton broth(supplemented with calcium and magnesium, or with sodium chloride foroxacillin, or with thymidine phosphorylase for trimethoprim,sulfamethoxazole and trimethoprim/sulfamethoxazole) to concentrationsbridging the range of clinical interest. One well on the 96-wellMicroscan® plate is a growth control well that contains dehydrated brothonly. The remaining wells contain dehydrated broth and antibiotic (orbroth and biochemical reagent indicator), which is rehydrated to thedesired concentration by inoculation of a standardized suspension oftest organism. The chromogenic biochemical agent indicators are used toidentify and characterize the species of bacteria based on detection ofpH changes and substrate utilization. After incubation overnight, theminimum inhibitory concentration (MIC) of an antibiotic for the testorganism is determined by observing the well with the lowestconcentration of the antibiotic that shows inhibition of growth. Twotypes of panel plates were utilized to test these gram-positiveorganisms: the Pos Combo Panel Type 6, and the Pos MIC Panel Type 6(both available from Baxter Diagnostics, Inc., Deerfield, Ill.). Theconcentrations of antibiotics tested in each panel are shown in Tables 2and 3 below.

                                      TABLE 2                                     __________________________________________________________________________    Antibiotic Concentrations (μg/mL) Tested in Pos Combo Panel Type 6         Plate                                                                         __________________________________________________________________________    Trimethoprim/Sulfamethoxazole                                                               2/38                                                            Rifampin      2   1                                                           Imipenem      8   4                                                           Cephalothin   16  8                                                           Amoxicillin/K Clavulanate                                                                   16/8                                                                              8/4                                                                              4/2                                                      Cefotaxime    32  8                                                           Ciprofloxacin 2   1                                                           Norfloxacin   8   4                                                           Nitrofurantoin                                                                              64  32                                                          Gentamicin    6   4  2  1                                                     Clindamycin   2   1  0.5                                                                              0.25                                                  Cefazolin     16  8  4  2                                                     Erythromycin  4   2  1  0.5                                                                              0.25                                               Vancomycin    16  8  4  2                                                     Penicillin    8   4  2  1  0.5                                                                              0.25                                                                             0.12                                                                             0.06                                                                             0.03                                   Ampicillin    8   4  2  1  0.5                                                                              0.25                                                                             0.12                                         Oxacillin     4   2  1  0.5                                                   Tetracycline  128 8  4  2                                                     __________________________________________________________________________

                                      TABLE 3                                     __________________________________________________________________________    Antibiotic Concentrations (μg/mL) Tested in Pos MIC Panel Type 6           __________________________________________________________________________    Plate                                                                         Erythromycin  4   2  1  0.5                                                                              0.25                                               Vancomycin    16  8  4  2                                                     Clindamycin   2   1  0.5                                                                              0.25                                                  Cefazolin     16  8  4  2                                                     Cephalothin   16  8  4  2                                                     Cefuroxime    16  8  4  2                                                     Ceftriaxone   32  16 8  4                                                     Chloramphenicol                                                                             16  8  4                                                        Cefotaxime    32  16 8  4                                                     Ticarcillin/K Clavulanate                                                                   8   4  2  1                                                     Imipenem      8   4  2  1                                                     Gentamicin    6   4  2  1                                                     Amoxicillin/K Clavulante                                                                    16/8                                                                              8/4                                                                              4/2                                                      Ampicillin/Sulbactam                                                                        16/8                                                                              8/4                                                         Rifampin      2   1                                                           Triethoprim/Sulfamethoxazole                                                                2/38                                                            Norfloxacin   8   4                                                           Penicillin    8   4  2  1  0.5                                                                              0.25                                                                             0.12                                                                             0.06                                                                             0.03                                   Ampicillin    8   4  2  1  0.5                                                                              0.25                                                                             0.12                                         Oxacillin     4   2  1  0.5                                                   Tetracycline  128 8  4  2                                                     Nitrofurantoin                                                                              64  32                                                          Ciprofloxacin 2   1                                                           Amikacin      32  16                                                          Sulfamethoxazole                                                                            256                                                             __________________________________________________________________________

For each experimental run, the following procedure was performed: Theorganism was streaked onto 5% sheep blood agar plates (Remel, Lenexa,Kans.) and incubated for 18-24 hours overnight. Well-isolated coloniesfrom the plates were emulsified in 3 ml of sterile Inoculum Water(catalog no. B1015-2, MicroScan® system, Baxter Diagnostics, Inc.,Deerfield, Ill.) to a final turbidity equivalent to 0.5 McFarland BariumSulfate standard. This cell suspension was vortexed for 2 to 3 secondsand 100 μl was transferred to glass tubes containing 25 ml of InoculumWater with Pluronic-D (catalog no. B1015-7, MicroScan® system, BaxterDiagnostics, Inc., Deerfield, Ill.) (hereinafter "Pluronic InoculumWater"), or 25 ml of Pluronic Inoculum Water into which the BPI proteinproduct (in formulation buffer) had been diluted to the desiredconcentration, generally between 0 and 64 μg/mL.

The 25 ml of this inoculum containing the BPI protein product was mixedby inversion and poured into a tray. The inoculum was drawn up into amanual 96-well pipetting system (RENOK® rehydrator-inoculator system,Baxter Health Care Corporation, West Sacramento, Calif.) designed foruse with the Microscan® panel plates, and 110 μl of the inoculum wasdelivered to each well of a Microscan® Pos Combo Panel Type 6 or Pos MICPanel Type 6 plate. When added to the wells, this inoculum achieves afinal bacterial concentration of 4×10⁵ to 7×10⁵ CFU/ml. The panel plateswere then incubated at 35° C. for 15-24 hours and read visually for cellgrowth.

No growth was defined as a slight whiteness in the well or a clearbroth. Growth appeared as turbidity which could take the form of a whitehaze throughout the well, a white button in the center of the well, or afine granule growth throughout the well. All wells were read against ablack indirectly lighted background. Visual results of the biochemicalreactions were read into a database for bacterial identification. TheMICs for each antibiotic tested were determined by identifying thelowest concentration of antibiotic which inhibited visible growth.

Table 4 below displays a summary of the results of the antibioticscreening panels, reported for each strain tested as the MIC of thetested antibiotics at varying concentrations of rBPI₂₁. Results are onlyreported where rBPI₂₁ altered the antibiotic susceptibility. Theantibiotic susceptibility standards (interpretation of an MIC asclinically resistant, intermediate or susceptible according to NCCLSstandards) applicable to the organism tested appear in Table 4A. Starsafter the antibiotic name in the "antibiotic tested" column indicatewhether rBPI₂₁ reversed the resistance of that organism to theantibiotic tested (two stars) or converted an indifferent MIC into asusceptible MIC (one star). These data show that rBPI₂₁ converted oneampicillin-resistant strain of S. pneumoniae into ampicillin-susceptible(the other tested strains were already sensitive to ampicillin) andincreased the susceptibility of other S. pneumoniae strains to amikacin,ampicillin, gentamicin and penicillin.

Table 4 also shows the presence or absence of bacterial growth in thegrowth control wells, which contained varying concentrations of rBPI₂₁alone without antibiotic. A "G" signifies growth at the testedconcentration, while "NG" signifies no growth. These results indicatethat S. pneumoniae is a BPI-susceptible organism; rBPI₂₁ was directlybactericidal/growth inhibitory for all tested isolates of S. pneumoniaeat a concentration of 2 μg/ml.

Additional screening of S. pneumoniae strains was conducted with avariety of BPI protein products, rBPI₂₁, rBPI₂₃, rBPI₅₀ and rBPI₄₂dimer. Table 4X below displays a summary of the results of theantibiotic screening panels, reported for each strain tested as the MICof the tested antibiotics at varying concentrations of the indicated BPIprotein product. Results are only reported if antibiotic susceptibilitywas altered.

                  TABLE 4                                                         ______________________________________                                        EFFECTS OF rBPI.sub.21 ± ANTITBIOTICS ON Streptococcus                     ______________________________________                                        pneumoniae                                                                               Minimum Inhibitory Concentration of                                           Antibiotic (μg/mL)                                                               With    With  With  With  With                               Microscan        0       0.5   2     8     16                                 Library                                                                              Antibiotic                                                                              μg/mL                                                                              μg/mL                                                                            μg/mL                                                                            μg/mL                                                                            μg/mL                           ID No. Tested    rBPI.sub.21                                                                           rBPI.sub.21                                                                         rBPI.sub.21                                                                         rBPI.sub.21                                                                         rBPI.sub.21                        ______________________________________                                        31573  --        G       G     NG    NG    NG                                        Ampicillin**                                                                            8       <0.12 <0.12 <0.12 <0.12                                     Penicillin*                                                                             0.5     <0.03 <0.03 <0.03 <0.03                                     Gentamicin*                                                                             6       2     <1    <1    <1                                 31582  --        G       --    NG    NG    NG                                        Ampicillin*                                                                             0.5     --    <0.12 <0.12 <0.12                                     Penicillin                                                                              0.06    --    <0.03 <0.03 <0.03                                     Gentamicin                                                                              <1      --    <1    <1    <1                                 15972  --        G       G     NG    NG    NG                                        Ampicillin                                                                              <0.12   <0.12 <0.12 <0.12 <0.12                                     Penicillin                                                                              <0.03   <0.03 <0.03 <0.03 <0.03                                     Gentamicin                                                                              4       2     <1    <1    <1                                        Amikacin* 32      --    --    --    <16                                015-035                                                                              --        G       G     NG    NG    NG                                        Ampicillin                                                                              <0.12   <0.12 <0.12 <0.12 <0.12                                     Peniciliin                                                                              <0.03   <0.03 <0.03 <0.03 <0.03                                     Gentamicin                                                                              4       <1    <1    <1    <1                                 015-034                                                                              --        G       --    NG    NG    NG                                        Ampicillin                                                                              <0.12   --    <0.12 <0.12 <0.12                                     Penicillin                                                                              <0.03   --    <0.03 <0.03 <0.03                                     Gentamicin                                                                              <1      --    <1    <1    <1                                 ______________________________________                                                         Minimum Inhibitory                                                            Concentration of                                                              Antibiotic (μg/mL)                                                                               With                                   Name                   With     With   16                                     (Microscan ®                                                                       Antibiotic    0 μg/mL                                                                             4 μg/mL                                                                           μg/mL                               ID No.   Tested        rBPI.sub.21                                                                            rBPI.sub.21                                                                          rBPI.sub.21                            ______________________________________                                        S. pneumoniae                                                                          --            G        NG     NG                                     1293     Erythromycin* 1        <.25   <.25                                            Clindamycin** >2       <.25   <.25                                            Cefuroxime    4        <2     <2                                              Penicillin*   1        <.03   <.03                                            Chloramphenicol*                                                                            16       <4     <4                                              Gentamicin    4        <1     <1                                              Ampicillin    1        <.12   <.12                                            Norfloxacin** >8       <4     <4                                              Ciprofloxacin*                                                                              2        <1     <1                                              Amikacin*     32       <16    <16                                    S. pneumoniae                                                                          --            G        NG     NG                                     130      Penicillin    .06      <.03   <.03                                            Gentamicin*   6        <1     <1                                              Trimethoprim/ >2       <2     <2                                              Sulfamethoxazole**                                                            Amikacin**    >32      <16    <16                                    S. pneumoniae                                                                          --            G        NG     NG                                     145      Cephalothin   4        <2     <2                                              Penicillin*   0.5      <.03   <.03                                            Gentamicin    4        <1     <1                                              Ampicillin    1        <.12   <.12                                            Trimethoprim/ >2       <2     <2                                              Sulfamethoxazole**                                                   S. pneumoniae                                                                          --            G        NG     NG                                     152      Gentamicin    4        <1     <1                                              Amikacin*     32       <16    <16                                    S. pneumoniae                                                                          --            G        G      NG                                     154      Erythromycin* 1        <.25   <.25                                            Cefuroxime    4        <2     <2                                              Penicillin*   1        .12    <.03                                            Chloramphenicol*                                                                            16       8      <4                                              Gentamicin    4        <1     <1                                              Ampicillin    2        0.5    <.12                                   ______________________________________                                         *BPI protein product reversed antibiotic indifference                         **BPI protein product reversed antibiotic resistance                     

                  TABLE 4A                                                        ______________________________________                                        Susceptibility ranges for S. pneumoniae                                               MIC (μg/mL)                                                        Antibiotic                                                                              Resistant   Intermediate                                                                            Susceptible                                   ______________________________________                                        Amikacin  >32         32        ≦16                                    Ampicillin                                                                              ≧4   0.25-2    ≦0.12                                  Gentamicin                                                                              >6           6        ≦4                                     Penicillin                                                                              ≧2   0.12-1    ≦0.06                                  ______________________________________                                    

                                      TABLE 4X                                    __________________________________________________________________________    EFFECTS OF BPI PROTEIN PRODUCTS ± ANTIBIOTICS                                              Minimum Inhibitory Concentration of Antibiotic (μg/mL)                     With:                                                         Organism                        4               16                            Name            0   4   4   4   μg/mL                                                                          16  16  16  μg/mL                      (Microscan ®                                                                     Antibiotic                                                                             μg/mL                                                                          μg/mL                                                                          μg/mL                                                                          μg/mL                                                                          rBPI.sub.42                                                                       μg/mL                                                                          μg/mL                                                                          μg/mL                                                                          rBPI.sub.42                   ID No.)                                                                              Tested   BPI rBPI.sub.23                                                                       rBPI.sub.21                                                                       BPI.sub.50                                                                        dimer                                                                             rBPI.sub.23                                                                       rBPI.sub.21                                                                       rBPI.sub.50                                                                       dimer                         __________________________________________________________________________    S.     --       G   G   NG  G   G   G   NG  G   NG                            pneumoniae                                                                           Erythromycin*                                                                          1   <0.25                                                                             <0.25                                                                             <0.25                                                                             <0.25                                                                             <0.25                                                                             <0.25                                                                             <0.25                                                                             <0.25                         1293   Clindamycin**                                                                          >2  <0.25                                                                             <0.25                                                                             <0.25                                                                             <0.25                                                                             <0.25                                                                             <0.25                                                                             <0.25                                                                             <0.25                                Cefuroxime                                                                             4   <2  <2  4   <2  <2  <2  <2                                       Penicillin*                                                                            1   0.12                                                                              <0.03                                                                             0.5 <0.03                                                                             0.5 <0.03                                                                             0.5 <0.03                                Chloramphenicol                                                                        16  <4  <4  8   <4  8   <4  <4  <4                                   Gentamicin                                                                             4   <1  <1  2   <1  4   <1  2   <1                                   Ampicillin                                                                             1   <0.12                                                                             <0.12                                                                             2   0.5 1   <0.12                                                                             1   1                                    Norfloxacin                                                                            >8  <4  <4  <4  <4  <4  <4  <4  <4                                   Trimethoprim/                                                                          >2  <2  <2  >2  <2  >2  <2  <2  <2                                   Sulfamethoxazole                                                              Ciprofloxacin                                                                          2   <1  <1  2   <1  <1  <1  <1  <1                                   Amikacin 32  <16 <16 <16 <16 <16 <16 <16 <16                           S.     --       G   NG  NG  G   NG  G   NG  NG  NG                            pneumoniae                                                                           Penicillin                                                                             0.06                                                                              <0.03                                                                             <0.03                                                                             <0.03                                                                             <0.03                                                                             <0.03                                                                             <0.03                                                                             <0.03                                                                             <0.03                         130    Gentamicin                                                                             6   <1  <1  6   <1  4   <1  <1  <1                                   Trimethoprim/                                                                          >2  <2  <2  <2  <2  >2  <2  <2  <2                                   Sulfamethoxazole                                                              Amikacin >32 <16 <16 >32 <16 32  <16 <16 <16                           S.     --       G   G   NG  G   NG  G   NG  G   NG                            pneumoniae                                                                           Cefazolin                                                                              <2  <2  <2  <2  <2  4   <2  <2  <2                            145    Cephalothin                                                                            4   <2  <2  <2  <2  4   <2  <2  <2                                   Penicillin*                                                                            0.5 0.06                                                                              <0.03                                                                             0.5 1   1   <0.03                                                                             <0.03                                                                             1                                    Gentamicin                                                                             4   <1  <1  2   <1  2   <1  <1  <1                                   Ampicillin                                                                             1   0.25                                                                              <0.12                                                                             0.5 0.5 2   <0.12                                                                             <0.12                                                                             2                                    Trimethoprim/                                                                          >2  <2  <2  <2  <2  >2  <2  <2  >2                                   Sulfamethoxazole                                                       S.     --       G   G   NG  G   G   G   NG  G   NG                            pneumoniae                                                                           Gentamicin                                                                             4   4   <1  <1  2   4   <1  <1  <1                            152    Amikacin*                                                                              32  <16 <16 32  <16 <16 <16 <16 <16                           S.     --       G   G   G   G   NG  G   NG  G   G                             pneumoniae                                                                           Erythromycin*                                                                          1   <0.25                                                                             <0.25                                                                             0.5 <0.25                                                                             <0.25                                                                             <0.25                                                                             1   <0.25                         154    Cefuroxime                                                                             4   <2  <2  4   <2  <2  <2  <2  <2                                   Penicillin*                                                                            1   0.25                                                                              0.12                                                                              1   <0.03                                                                             0.5 <0.03                                                                             0.5 0.25                                 Chloramphenicol*                                                                       16  8   8   8   <4  8   <4  8   <4                                   Gentamicin                                                                             4   2   <1  2   <1  4   <1  4   2                                    Ampicillin**                                                                           2   0.5 0.5 1   <0.12                                                                             2   <0.12                                                                             2   <0.12                                Amikacin 32  <16 <16 32  <16 <16 <16 <16 <16                           S.     Penicillin                                                                             0.12                                                                              <0.03                                                                             <0.03                                                                             0.12                                                                              0.06                                                                              0.06                                                                              0.06                                                                              0.12                                                                              <.03                          pneumoniae                                                                           Gentamicin                                                                             >6  >6  6   >6  >6  >6  >6  >6  6                             141    Trimethoprim/                                                                          >2  <2  <2  >2  <2  <2  <2  <2  <2                                   Sulfamethoxazole**                                                            Amikacin >32 >32 >32 >32 >32 >32 >32 >32 32                            S.     Cefazolin                                                                              4   <2  <2  4   <2  <2  <2  4   <2                            pneumoniae                                                                           Cephalothin                                                                            4   <2  <2  4   <2  4   <2  4   <2                            145    Penicillin*                                                                            0.5 <0.03                                                                             <0.03                                                                             0.5 <0.03                                                                             1   <0.03                                                                             0.5 <0.03                                Gentamicin                                                                             4   4   <1  4   <1  6   <1  6   <1                                   Ampicillin**                                                                           1   <0.12                                                                             <0.12                                                                             2   <0.12                                                                             2   <0.12                                                                             2   <0.12                                Trimethoprim/                                                                          >2  <2  <2  >2  <2  >2  <2  >2  <2                                   Sulfametoxazole**                                                             Amikacin*                                                                              32  <16 <16 <16 <16 32  <16 32  <16                           __________________________________________________________________________

EXAMPLE 10 In Vitro Effects of BPI Protein Product Alone or inCombination with Antibiotics on Strains of the Gram-Positive OrganismStreptococcus pyogenes (Group A)

The direct growth inhibitory effect of a BPI protein product, rBPI₂₁, onvarious strains of Streptococcus pyogenes, also known as Group A strep,was evaluated using the Microscan® screening assay of Example 9. Theeffect of rBPI₂₁ on the antibiotic susceptibility of these strains wasalso evaluated in the same assay. Assays were conducted on clinicalisolates of S. pyogenes (from Baxter Microscan® library, Sacramento,Calif.).

A summary of the results of the BPI protein product and antibioticscreening panels, reported as MICs (μg/ml) of the tested antibiotic atvarying concentrations of rBPI₂₁ (μg/ml), is shown in Table 5 below.Results are only reported where rBPI₂₁ altered the antibioticsusceptibility or growth. The antibiotic susceptibility standards(interpretation of an MIC as clinically resistant, intermediate orsusceptible according to NCCLS standards) applicable to the organismtested appear in Table 5A. Table 5 also shows the presence or absence ofbacterial growth ("G" or "NG") in the control well, which containsvarying concentrations of rBPI₂₁ alone without antibiotic.

These results indicate that rBPI₂₁ alone had a directbactericidal/growth inhibitory effect on one strain of S. pyogenes(Group A) at a concentration as low as 2 μg/ml, on three of the strainsat 8 μg/ml, and on the remaining strain at 32 μg/ml. The data also showthat BPI protein product increased the antibiotic susceptibility ofStrep. pyogenes (Group A) strains to norfloxacin and gentamicin.

                                      TABLE 5                                     __________________________________________________________________________    EFFECTS OF rBPI.sub.21 ± ANTIBIOTICS ON Streptococcus pyogenes (Group      A)                                                                                       Minimum inhibitory Concentration of Antibiotic (μg/mL)                     With                                                                              With          With                                                                              With                                         Microscan  0   0.5 With With 16  32                                           Library                                                                            Antibiotic                                                                          μg/mL                                                                          μg/mL                                                                          2 μg/mL                                                                         8 μg/mL                                                                         μg/mL                                                                          μg/mL                                     ID No.                                                                             Tested                                                                              rBPI.sub.21                                                                       rBPI.sub.21                                                                       rBPI.sub.21                                                                        rBPI.sub.21                                                                        rBPI.sub.21                                                                       rBPI.sub.21                                  __________________________________________________________________________    30847                                                                              --    G   G   G    NG   NG  NG                                                Norfloxacin                                                                         <4  <4  <4   <4   <4  <4                                                Gentamicin                                                                          2   2   <1   <1   <1  <1                                           002-002                                                                            --    G   G   G    G    G   NG                                                Norfloxacin                                                                         <4  <4  <4   <4   <4  <4                                                Gentamicin                                                                          2   <1  <1   <1   <1  <1                                           19035                                                                              --    G   G   G    NG   G   NG                                                Norfloxacin                                                                         <4  <4  <4   <4   <4  <4                                           30403                                                                              --    G   G   NG   NG   NG  NG                                                Norfloxacin*                                                                        8   <4  <4   <4   <4  <4                                           30413                                                                              --    G   G   G    NG   G   NG                                                Norfloxacin*                                                                        8   <4  <4   <4   <4  <4                                           __________________________________________________________________________     *BPI protein product reversed antibiotic indifference                    

                  TABLE 5A                                                        ______________________________________                                        Susceptibility ranges for S. pyogenes (Group A)                                       MIC (μg/mL)                                                        Antibiotic                                                                              Resistant   Intermediate                                                                            Susceptible                                   ______________________________________                                        Gentanmicin                                                                             >6          6         ≦4                                     Norfloxacin                                                                             >8          8         ≦4                                     ______________________________________                                    

EXAMPLE 11 In Vitro Effects of BPI Protein Product Alone or inCombination with Antibiotics on Strains of the Gram-Positive OrganismStreptococcus agalactiae (Group B)

The effect of a BPI protein product, rBPI₂₁, on the antibioticsusceptibility of various strains of Streptococcus agalactiae, alsoknown as Group B strep, was evaluated using the Microscan® antibioticsusceptibility screening assay of Example 9. The direct growthinhibitory effect of rBPI₂₁ on these strains was also evaluated in thesame assay. Assays were conducted on clinical isolates of S. agalactiae(from Baxter Microscan® library, Sacramento, Calif.).

A summary of the results of the antibiotic screening panels, reported asMICs (μg/ml) of the antibiotic tested, is shown in Table 6 below.Results are only reported where rBPI₂₁ altered the antibioticsusceptibility. The antibiotic susceptibility standards (interpretationof an MIC as clinically resistant, intermediate or susceptible accordingto NCCLS standards) applicable to the organism tested appear in Table6A. These results show that BPI protein product reversed the resistanceof one strain to gentamicin, and increased the susceptibility of otherstrains to ciprofloxacin, gentamicin, norfloxacin. These strains werenot susceptible to BPI protein product alone, without antibiotic.

                  TABLE 6                                                         ______________________________________                                        EFFECTS OF rBPI.sub.21 ± ANTIBIOTICS ON                                    Steptococcus agalactiae (Group B)                                                        Minimum Inhibitory Concentration of                                           Antibiotic (μg/mL)                                                               With    With  With  With  With                               Microscan        0       0.5   2     8     16                                 Library                                                                              Antibiotic                                                                              μg/mL                                                                              μg/mL                                                                            μg/mL                                                                            μg/mL                                                                            μg/mL                           ID No. Tested    rBPI.sub.21                                                                           rBPI.sub.21                                                                         rBPI.sub.21                                                                         rBPI.sub.21                                                                         rBPI.sub.21                        ______________________________________                                        003-044                                                                              Gentamicin*                                                                             >6      6     4     6     6                                         Ciprofloxa-                                                                             <1      <1    <1    <1    <1                                        cin                                                                           Norfloxacin                                                                             <4      <4    <4    <4    <4                                 003-049                                                                              Gentami-  >6      4     4     4     4                                         cin**                                                                         Ciproflox-                                                                              2       2     <1    <1    <1                                        acin*                                                                         Norfloxacin                                                                             >8      8     8     8     8                                         Gentamicin                                                                              4       <1    2     2     <1                                 30851  Ciprofloxa-                                                                             <1      <1    <1    <1    <1                                        cin                                                                           Norfloxacin*                                                                            8       <4    <4    <4    <4                                        Gentamicin                                                                              2       4     <1    <1    <1                                 31947  Ciproflox-                                                                              2       <1    <1    <1    <1                                        acin*                                                                         Norfloxacin                                                                             8       8     8     8     8                                         Gentamicin                                                                              4       <1    4     <1    <1                                 30911  Ciprofloxa-                                                                             <1      <1    <1    <1    <1                                        cin                                                                           Norfloxacin                                                                             <4      <4    <4    <4    <4                                 ______________________________________                                         *BPI protein product reversed antibiotic indifference                         **BPI protein product reversed antibiotic resistance                     

                  TABLE 6A                                                        ______________________________________                                        Susceptibility ranges for S. agalactiae (Group B)                                      MIC (μg/mL)                                                       Antibiotic Resistant   Intermediate                                                                            Susceptible                                  ______________________________________                                        Ciprofloxacin                                                                            >2          2         ≦1                                    Gentamicin >6          6         ≦4                                    Norfloxacin                                                                              >8          8         ≦4                                    ______________________________________                                    

EXAMPLE 12 In Vitro Effects of BPI Protein Products Alone or inCombination with Antibiotics on Strains of the Gram-Positive OrganismStreptococcus bovis

The effect of a BPI protein product, rBPI₂₁, on the antibioticsusceptibility of various strains of Streptococcus bovis was evaluatedusing the Microscan® antibiotic susceptibility screening assay ofExample 9. The direct growth inhibitory effect of rBPI₂₁ on thesestrains was also evaluated in the same assay. Assays were conducted onclinical isolates of S. bovis (from Baxter Microscan® library,Sacramento, Calif.).

A summary of the results of the antibiotic screening panels, reported asMICs (μg/ml) of the antibiotic tested, is shown in Table 7 below.Results are only reported where rBPI₂₁ altered the antibioticsusceptibility. The antibiotic susceptibility standards (interpretationof an MIC as clinically resistant, intermediate or susceptible accordingto NCCLS standards) applicable to the organism tested appear in Table7A. These results show that the BPI protein product reversed theresistance of one strain to ciprofloxacin, three strains to norfloxacin,and two strains to tetracycline. The results show that BPI proteinproduct also increased the susceptibility of some strains tociprofloxacin, gentamicin, norfloxacin, and tetracycline. These strainswere not susceptible to BPI protein product alone, without antibiotic.

                                      TABLE 7                                     __________________________________________________________________________    EFFECTS OF rBPI.sub.21 ± ANTIBIOTICS ON Streptococcus bovis                              Minimum Inhibitory Concentration of Antibiotic                                (μg/mL)                                                                    With With           With                                        Microscan     0    0.5 With  With 16                                          Library                                                                             Antibiotic                                                                            μg/mL                                                                           μg/mL                                                                          2 μg/mL                                                                          8 μg/mL                                                                         μg/mL                                    ID No.                                                                              Tested  rBPI.sub.21                                                                        rBPI.sub.21                                                                       rBPI.sub.21                                                                         rBPI.sub.21                                                                        rBPI.sub.21                                 __________________________________________________________________________    ATCC# Tetracycline*                                                                         128  8   8     4    8                                           49147 Gentamicin                                                                            2    2   <1    <1   <1                                                Ciprofloxacin                                                                         <1   <1  <1    <1   <1                                                Norfloxacin*                                                                          8    <4  <4    <4   <4                                          008-009                                                                             Tetracycline**                                                                        128  4   8     4    <2                                                Gentamicin                                                                            2    2   2     <1   <1                                                Ciprofloxacin*                                                                        2    <1  <1    <1   <1                                                Norfloxacin**                                                                         >8   8   8     8    <4                                          008-010                                                                             Tetracycline**                                                                        128  128 128   8    4                                                 Gentamicin                                                                            4    4   2     2    <1                                                Ciprofloxacin*                                                                        2    2   <1    <1   <1                                                Norfloxacin**                                                                         >8   8   >8    <4   <4                                          008-011                                                                             Tetracycline                                                                          <2   <2  <2    <2   <2                                                Gentamicin                                                                            2    2   <1    <1   <1                                                Ciprofloxacin*                                                                        2    <1  2     <1   <1                                                Norfloxacin**                                                                         >8   8   8     8    <4                                          008-012                                                                             Tetracycline                                                                          128  128 128   128  8                                                 Gentamicin                                                                            4    2   <1    2    <1                                                Ciprofloxacin**                                                                       >2   2   <1    2    <1                                                Norfloxacin                                                                           >8   8   8     8    8                                           __________________________________________________________________________     *BPI protein product reversed antibiotic indifference                         **BPI protein product reversed antibiotic resistance                     

                  TABLE 7A                                                        ______________________________________                                        Susceptibility ranges for S. bovis                                                     MIC (μg/mL)                                                       Antibiotic Resistant   Intermediate                                                                            Susceptible                                  ______________________________________                                        Ciprofloxacin                                                                            >2          2         ≦1                                    Gentamicin >6          6         ≦4                                    Norfloxacin                                                                              >8          8         ≦4                                    Tetracycline                                                                             ≧128 8         ≦4                                    ______________________________________                                    

EXAMPLE 13 In Vitro Effects of BPI Protein Products Alone or inCombination with Antibiotics on Strains of the Gram-Positive OrganismEnterococcus faecalis

The effect of a BPI protein product, rBPI₂₁, on the antibioticsusceptibility of various strains of Enterococcus faecalis was evaluatedusing the Microscan® antibiotic susceptibility screening assay ofExample 9. The direct growth inhibitory effect of rBPI₂₁ on thesestrains was also evaluated in the same assay. Assays were conducted onclinical isolates of E. faecalis (from Baxter Microscan® library,Sacramento, Calif.).

A summary of the results of the antibiotic screening panels, reported asMICs (μg/ml) of the antibiotic tested, is shown in Table 8 below.Results are only reported where rBPI₂₁ altered the antibioticsusceptibility. The antibiotic susceptibility standards (interpretationof an MIC as clinically resistant, intermediate or susceptible accordingto NCCLS standards) applicable to the organism tested appear in Table8A. These results show that BPI protein product reversed the resistanceof three strains to ciprofloxacin and two strains to norfloxacin. TheBPI protein product also increased the susceptibility of some E.faecalis strains to ampicillin, ciprofloxacin, erythromycin,norfloxacin, penicillin, rifampin, and tetracycline. These strains werenot susceptible to BPI protein product alone, without antibiotic.

                                      TABLE 8                                     __________________________________________________________________________    EFFECTS OF rBPI.sub.21 ± ANTIBIOTICS ON Enterococcus faecalis                             Minimum Inhibitory Concentration of Antibiotic (μg/mL)      Microscan      With  With  With  With                                         Library                                                                             Antibiotic                                                                             0 μg/ml                                                                          2 μg/mL                                                                          8 μg/ml                                                                          32 μg/mL                                  ID. No.                                                                             Tested   rBPI.sup.21                                                                         rBPI.sub.21                                                                         rBPI.sub.21                                                                         rBPI.sub.21                                  __________________________________________________________________________    011-070                                                                             Penicillin                                                                             2     2     1     1                                                  Tetracycline                                                                           128   128   128   8                                            16967 Rifampin >2    >2    >2    2                                                  Ciprofloxacin**                                                                        >2    2     2     <1                                                 Norfloxacin**                                                                          >8    8     8     <4                                                 Penicillin                                                                             2     2     2     1                                                  Erythromycin                                                                           4     2     2     2                                            19010 Penicillin                                                                             8     8     8     4                                                  Ampicillin                                                                             4     4     4     2                                            011-028                                                                             Penicillin                                                                             2     2     2     1                                                  Ciprofloxacin*                                                                         2     <1    <1    <1                                                 Ampicillin                                                                             1     <0.5  <0.5  <0.5                                         011-001                                                                             Ciprofloxacin**                                                                        >2    <1    <1    <1                                                 Norfloxacin                                                                            <4    <4    <4    <4                                           011-002                                                                             Penicillin                                                                             2     1     1     1                                                  Ampicillin                                                                             2     0.5   0.5   0.5                                          011-003                                                                             Rifampin >2    >2    >2    2                                                  Penicillin                                                                             2     2     2     1                                                  Norfloxacin*                                                                           8     8     8     <4                                                 Ampicillin                                                                             1     1     0.5   0.5                                          011-004                                                                             Penicillin                                                                             8     8     8     4                                                  Ampicillin                                                                             4     4     2     2                                            011-005                                                                             Penicillin                                                                             2     2     2     1                                                  Ampicillin                                                                             1     0.5   0.5   0.5                                          011-006                                                                             Penicillin                                                                             2     2     2     1                                                  Ampicillin                                                                             1     1     1     0.5                                          011-066                                                                             Ciprofloxacin**                                                                        >2    <1    <1    <1                                                 Norfloxacin**                                                                          >8    <4    <4    <4                                                 Ampicillin                                                                             1     1     0.5   0.5                                          __________________________________________________________________________     *BPI protein product reversed antibiotic indifference                         **BPI protein product reversed antibiotic resistance                     

                  TABLE 8A                                                        ______________________________________                                        Susceptibility ranges for E. faecalis                                                  MIC (μg/mL)                                                       Antibiotic Resistant  Intermediate                                                                            Susceptible                                   ______________________________________                                        Ampicillin >8                   ≦8                                     Ciprofloxacin                                                                            >2         2         ≦1                                     Erythromycin                                                                             >4         1-4       ≦0.5                                   Norfloxacin                                                                              >8         8         ≦4                                     Penicillin >8                   ≦8                                     Rifampin   >2         2         ≦1                                     Tetracycline                                                                             >128       8         ≦4                                     ______________________________________                                    

EXAMPLE 14 In Vitro Effects of BPI Protein Products Alone or inCombination with Antibiotics on Strains of the Gram-Positive OrganismEnterococcus faecium

The effect of a variety of BPI protein products on the antibioticsusceptibility of various strains of Enterococcus faecium was evaluatedusing the Microscan® antibiotic susceptibility screening assay ofExample 9. The direct growth inhibitory effect of the BPI proteinproducts on these strains was also evaluated in the same assay. Assayswere conducted on clinical isolates of E. faecium (from BaxterMicroscan® library, Sacramento, Calif.).

A summary of the results of the antibiotic screening panels, reported asMICs (μg/ml) of the antibiotic tested at varying concentrations ofrBPI₂₁, is shown in Table 9 below. Results are only reported whererBPI₂₁ altered the antibiotic susceptibility. The antibioticsusceptibility standards (interpretation of an MIC as clinicallyresistant, intermediate or susceptible according to NCCLS standards)applicable to the organism tested appear in Table 9A. These results showthat rBPI₂₁ reversed the resistance of five strains to amoxicillin/Kclavulanate, two strains to cefazolin, two strains to ciprofloxacin, twostrains to erythromycin, one strain to norfloxacin, two strains tooxacillin, two strains to rifampin, one strain to tetracycline, and onestrain to vancomycin. The rBPI₂₁ also increased the susceptibility ofsome E. faecium strains to amoxicillin/K clavulanate, cefotaxime,ciprofloxacin, erythromycin, penicillin, rifampin, and vancomycin. Thesestrains were not susceptible to BPI protein product alone, withoutantibiotic.

Additional screening of an E. faecium strain (Microscan® ID no. 15773)was conducted with a variety of BPI protein products, rBPI₂₁, rBPI₂₃,rBPI₅₀ and rBPI₄₂ dimer. Table 9X below displays a summary of theresults of the antibiotic screening panels, reported as the MIC of thetested antibiotics at varying concentrations of the indicated BPIprotein product. Results are only reported if antibiotic susceptibilitywas altered.

                                      TABLE 9                                     __________________________________________________________________________    EFFECTS OF rBPI.sub.21 ± ANTIBIOTICS ON Enterococcus faecium                              Minimum Inhibitory Concentration of Antibiotic (μg/mL)      Microscan      With  With  With  With                                         Library                                                                             Antibiotic                                                                             0 μg/mL                                                                          2 μg/mL                                                                          8 μg/mL                                                                          32 μg/mL                                  ID. No.                                                                             Tested   rBPI.sup.21                                                                         rBPI.sub.21                                                                         rBPI.sub.21                                                                         rBPI.sub.21                                  __________________________________________________________________________    17611 Rifampin**                                                                             >2    >2    2     <1                                                 Ciprofloxacin**                                                                        >2    2     2     <1                                                 Augmentin.sup.a                                                                        >16   16    16    16                                           16866 Ciprofloxacin*                                                                         2     2     <1    <1                                                 Tetracycline**                                                                         128   128   128   4                                            20076 Rifampin**                                                                             >2    2     2     <1                                                 Erythromycin*                                                                          4     2     1     <0.25                                              Augmentin.sup.a **                                                                     16    8     8     <4                                                 Oxacillin**                                                                            >4    <0.5  <0.5  <0.5                                         012-048                                                                             Rifampin >2    >2    2     2                                                  Ciprofloxacin**                                                                        >2    >2    <1    <1                                                 Penicillin                                                                             0.25  0.25  0.25  0.12                                               Cefotaxime*                                                                            32    <8    <8    <8                                                 Cefazolin**                                                                            >16   16    8     8                                            15773 Rifampin*                                                                              2     2     <1    <1                                                 Ciprofloxacin*                                                                         2     <1    <1    <1                                                 Erythromycin**                                                                         >4    <0.25 <0.25 <0.25                                              Augmentin.sup.a **                                                                     16    <4    <4    <4                                                 Vancomycin                                                                             4     4     <2    <2                                           012-001                                                                             Rifampin >2    2     >2    2                                                  Augmentin.sup.a **                                                                     16    8     8     8                                            012-002                                                                             Rifampin >2    >2    >2    2                                                  Oxacillin**                                                                            >4    1     <0.5  <0.5                                               Vancomycin**                                                                           >16   >16   >16   <2                                                 Norfloxacin**                                                                          >8    <4    <4    <4                                                 Cefazolin**                                                                            >16   >16   >16   8                                            012-003                                                                             Rifampin <1    <1    <1    <1                                                 Augmentin.sup.a **                                                                     16    8     8     8                                            012-004                                                                             Rifampin*                                                                              2     2     <1    <1                                                 Ciprofloxacin*                                                                         2     <1    <1    <1                                                 Augmentin.sup.a **                                                                     16    <4    <4    <4                                                 Vancomycin                                                                             4     4     <2    <2                                                 Erythromycin**                                                                         >4    0.5   <0.25 <0.25                                        __________________________________________________________________________     .sup.a Augmentin is Amoxicillin K/Clavulanate                                 *BPI protein product reversed antibiotic indifference                         **BPI protein product reversed antibiotic resistance                     

                  TABLE 9A                                                        ______________________________________                                        Susceptibility ranges for E. faecium                                                   MIC (μg/ml)                                                       Antibiotic Resistant  Intermediate                                                                            Susceptible                                   ______________________________________                                        Amoxicillin                                                                              ≧16/8          ≦8/4                                  K/Clavulanate                                                                 Cefazolin  >16        16        ≦8                                     Cefotaxime >32        16-32     ≦8                                     Ciprofloxacin                                                                            >2         2         ≦1                                     Erythromycin                                                                             >4         1-4         ≦0.5                                 Norfloxacin                                                                              >8         8         ≦4                                     Oxacillin  ≧4            ≦2                                     (staph only)                                                                  Penicillin >8                   ≦8                                     Rifampin   >2         2         ≦1                                     Tetracycline                                                                             ≧128                                                                              8         ≦4                                     Vancomycin >16         8-16     ≦4                                     ______________________________________                                    

                                      TABLE 9X                                    __________________________________________________________________________    EFFECTS OF BPI PROTEIN PRODUCTS ± ANTIBIOTICS                                              Minimum Inhibitory Concentration of Antibiotic (μg/mL)                     With:                                                         Organism                        4               16                            Name            0   4   4   4   μg/mL                                                                          16  16  16  μg/mL                      (Microscan ®                                                                     Antibiotic                                                                             μg/mL                                                                          μg/mL                                                                          μg/mL                                                                          μg/mL                                                                          rBPI.sub.42                                                                       μg/mL                                                                          μg/mL                                                                          μg/mL                                                                          rBPI.sub.42                   ID No.)                                                                              Tested   BPI rBPI.sub.21                                                                       rBPI.sub.23                                                                       rBPI.sub.50                                                                       dimer                                                                             rBPI.sub.21                                                                       rBPI.sub.23                                                                       rBPI.sub.50                                                                       dimer                         __________________________________________________________________________    E. faecium                                                                           Erythromycin**                                                                         >4  1   <0.25                                                                             >4  >4  <0.25                                                                             <0.25                                                                             >4  <0.25                         15773  Vancomycin                                                                             4   4   4   4   4   <2  4   4   <2                                   Cefotaxime                                                                             >32 >32 >32 >32 >32 >32 32  >32 >32                                  Ticarcillin/K                                                                          >8  >8  >8  >8  >8  >8  8   >8  >8                                   Clavulanate                                                                   Gentamicin**                                                                           >6  4   4   >6  >6  6   6   >6  4                                    Amoxicillin/K                                                                          16  8   <4  16  8   <4  <4  8   <4                                   Clavulanate                                                                   Ampicillin/                                                                            16  <8  <8  16  <8  <8  <8  <8  <8                                   Sulbactam                                                                     Rifampin >2  2   2   >2  2   <1  <1  >2  <1                                   Ciprofloxacin*                                                                         2   <1  <1  2   <1  <1  <1  <1  <1                                   Amikacin*                                                                              32  <16 <16 32  32  <16 32  32  <16                                  Sulfamethoxazole**                                                                     >256                                                                              <256                                                                              <256                                                                              >256                                                                              >256                                                                              <256                                                                              >256                                                                              >256                                                                              <256                          __________________________________________________________________________

EXAMPLE 15 In Vitro Effects of BPI Protein Products Alone or inCombination with Antibiotics on Other Gram-Positive Enterococcus Species

The effect of a BPI protein product, rBPI₂₁, on the antibioticsusceptibility of a variety of Enterococcus species was evaluated usingthe Microscan® antibiotic susceptibility screening assay of Example 9.The direct growth inhibitory effect of rBPI₂₁ on these species was alsoevaluated in the same assay. Assays were conducted on clinical isolatesof Enterococcus species (from Baxter Microscan® library, Sacramento,Calif.).

A summary of the results of the antibiotic screening panels, reported asMICs (μg/ml) of the antibiotic tested, is shown in Table 10 below.Results are only reported where rBPI₂₁ altered the antibioticsusceptibility. The antibiotic susceptibility standards (interpretationof an MIC as clinically resistant, intermediate or susceptible accordingto NCCLS standards) applicable to the organism tested appear in Table10A. These results show that for E. gallinarum, BPI protein productreversed resistance to cephalothin, cefazolin, ciprofloxacin andnorfloxacin, and increased susceptibility to ampicillin, penicillin andrifampin. For E. raffinosus, BPI protein product reversed resistance toampicillin and increased susceptibility to imipenem. For E.casseliflavus, BPI protein product increased susceptibility toampicillin, penicillin, oxacillin, cephalothin, and cefazolin. For E.durans, BPI protein product increased susceptibility to ampicillin andrifampin. For E. avium, BPI protein product increased susceptibility toampicillin, penicillin, cefotaxime and ciprofloxacin, and oxacillin.These Enterococcus species were not susceptible to BPI protein productalone, without antibiotic.

                                      TABLE 10                                    __________________________________________________________________________    EFFECTS OF rBPI.sub.21 ± ANTIBIOTICS ON Enterococcus SPECIES                                Minimum Inhibitory Concentration of                                           Antibiotic (μg/mL)                                                         With With With                                               Microscan        0    2    8    With                                          Library Antibiotic                                                                             μg/mL                                                                           μg/mL                                                                           μg/mL                                                                           32 μg/mL                                   ID No.  Tested   rBPI.sup.21                                                                        rBPI.sub.21                                                                        rBPI.sub.21                                                                        rBPI.sub.21                                   __________________________________________________________________________    031-009 Ampicillin                                                                             1    1    0.5  0.5                                           ( E. gallinarum)                                                                      Penicillin                                                                             1    0.5  0.5  0.25                                                  Rifampin >2   >2   >2   2                                                     Cephalothin**                                                                          >16  <8   <8   <8                                                    Cefazolin**                                                                            >16  8    8    4                                                     Ciprofloxacin**                                                                        >2   2    <1   <1                                                    Norfloxacin**                                                                          >8   8    8    <4                                            16206   Ampicillin**                                                                           >8   >8   >8   8                                             (E. raffinosus)                                                                       Imipenem >8   >8   >8   8                                             030-008 and                                                                           Ampicillin                                                                             1    0.5  0.25 0.25                                          030-001*                                                                              Penicillin                                                                             1    0.25 0.25 0.12                                          (E. casseliflavus)                                                                    Oxacillin                                                                              >4   >4   >4   4                                                     Cephalothin*                                                                           16   <8   <8   <8                                                    Cefazolin*                                                                             16   8    4    4                                             17543   Ampicillin                                                                             1    0.5  0.5  0.5                                           (E. durans)                                                                           Rifampin >2   2    2    2                                             31413   Ampicillin                                                                             0.5  0.5  0.5  <0.12                                         (E. avium)                                                                            Penicillin                                                                             1    1    1    0.5                                                   Oxacillin**                                                                            >4   >4   4    0.5                                                   Cefotaxime*                                                                            32   <8   <8   <88                                                   Ciprofloxacin*                                                                         2    <1   <1   <1                                            __________________________________________________________________________     .sup.a Two strains of E. casseliflavus were tested; both strains gave         identical results.                                                       

                  TABLE 10A                                                       ______________________________________                                        Susceptibility ranges for Enterococcus species                                         MIC (μg/mL)                                                       Antibiotic Resistant   Intermediate                                                                            Susceptible                                  ______________________________________                                        Ampicillin >8                    ≦8                                    Cefazolin  >16         16        ≦8                                    Cefotaxime >32         16-32     ≦8                                    Cephalothin                                                                              >16         16        ≦8                                    Ciprofloxacin                                                                            >2          2         ≦1                                    Imipenem   >8          8         ≦4                                    Norfloxacin                                                                              >8          8         ≦4                                    Oxacillin  ≧4             ≦2                                    Penicillin >8                    ≦8                                    Rifampin   >2          2         ≦1                                    ______________________________________                                    

EXAMPLE 16 In Vitro Effects of BPI Protein Products Alone or inCombination with Antibiotics on Strains of the Gram-Positive OrganismStaphylococcus aureus

The effect of BPI protein products on the antibiotic susceptibility of avariety of S. aureus species was evaluated using the Microscan®antibiotic susceptibility screening assay of Example 9. The directgrowth inhibitory effect of BPI protein products on these species wasalso evaluated in the same assay. Assays were conducted on the followingclinical isolates of S. aureus (from Baxter Microscan® library,Sacramento, Calif.): 052-066, 052-106, 052-107, 052-108, 052-184,052-219, 052-230, 14288, 20720, 29213 (ATCC No.), 32075 and 32073.

These strains of S. aureus were tested to determine the MIC₉₀ for eachantibiotic in the panel at varying concentrations of rBPI₂₁. The MIC₉₀is defined as the lowest concentration of antibiotic that inhibits thegrowth of 90% of all S. aureus isolates tested. Results are reported inTable 11 below, and applicable antibiotic susceptibility standards(interpretation of an MIC as clinically resistant, intermediate orsusceptible according to NCCLS standards) appear in Table 11A.

The data show that rBPI₂₁ was able to reduce the S. aureus MIC₉₀ valuesfor eight of twenty-five antibiotics evaluated: amoxicillin/Kclavulanate (Augmentin), cefotaxime, ceftriaxone, cefuroxime,cephalothin, chloramphenicol, imipenem and sulfamethoxazole. Dashes inTable 11 indicate that no concentration of antibiotic tested was able toinhibit growth of 90% of all tested strains. None of the S. aureusstrains tested were susceptible to rBPI₂₁ alone without antibiotic.

Comparable results obtained from testing of the BPI protein productrBPI₂₁ on S. aureus (Microscan® ID no. 052-106) in the Pasco systemconfirm the effect of BPI protein products on the antibioticsusceptibility of gram-positive organisms.

Additional screening of this S. aureus strain (Microscan® ID no.052-106) was conducted with a variety of BPI protein products, rBPI₂₁,rBPI₂₃, rBPI₅₀ and rBPI₄₂ dimer. Table 11X below displays a summary ofthe results of the antibiotic screening panels, reported as the MIC ofthe tested antibiotics at varying concentrations of the indicated BPIprotein product. Results are only reported if antibiotic susceptibilitywas altered.

                  TABLE 11                                                        ______________________________________                                        Effect of rBPI.sub.21 on MIC.sub.90 of Antibiotics                            for Staphylococcus aureus                                                                MIC.sub.90 With                                                                          MIC.sub.90 With                                                                          MIC.sub.90 With                                         0 μg/mL 0.5-8 μg/mL                                                                           16-64 μg/mL                                          rBPI.sub.21                                                                              rBPI.sub.21                                                                              rBPI.sub.21                                  Antibiotic Tested                                                                        (n = 38).sup.a                                                                           (n = 31).sup.a                                                                           (n = 25).sup.a                               ______________________________________                                        Amikacin   16         16         16                                           Amox/K Clav                                                                              16         16         8                                            Amp/Sulbactam                                                                            8          8          8                                            Ampicillin --         --         --                                           Cefazolin  --         --         --                                           Cefotaxime 32         8          4                                            Ceftriaxone                                                                              --         32         8                                            Cefuroxime --         --         16                                           Cephalothin                                                                              8          4          4                                            Chloramphenicol                                                                          8          8          4                                            Ciprofloxacin                                                                            1          1          1                                            Clindamycin                                                                              --         --         --                                           Erythromycin                                                                             --         --         --                                           Gentamicin --         --         --                                           Imipenem   --         --         8                                            Nitrofurantoin                                                                           32         32         32                                           Norfloxacin                                                                              4          4          4                                            Oxacillin  --         --         --                                           Penicillin --         --         --                                           Rifampin   1          1          1                                            Sulfamethoxazole                                                                         --         --         256                                          Tetracycline                                                                             128        128        128                                          Ticarcillin/K Clav                                                                       --         --         --                                           Trimeth/Sulfa                                                                            2          2          2                                            Vancomycin 2          2          2                                            ______________________________________                                         .sup.a Number of runs; 12 strains tested                                 

                  TABLE 11A                                                       ______________________________________                                        Susceptibility ranges for S. aureus                                                      MIC (μg/mL)                                                     Antibiotic   Resistant Intermediate                                                                             Susceptible                                 ______________________________________                                        Amikacin     >32       32          ≦16                                 Amoxicillin/K Clav.                                                                        ≧8/4           ≦4/2                                Ampicillin/Sulbactam                                                                       >16/8     16/8        ≦8/4                                Ampicillin     ≧0.5          ≦0.25                              Cefazolin    >16       16         ≦8                                   Cefotaxime   >32       16-32      ≦8                                   Ceftriaxone  >32       16-32      ≦8                                   Cefuroxime   >16       16         ≦8                                   Cephalothin  >16       16         ≦8                                   Chloramphenicol                                                                            >16       16         ≦8                                   Ciprofloxacin                                                                              >2        2          ≦1                                   Clindamycin  >2        1-2          ≦0.5                               Erythromycin >4        1-4          ≦0.5                               Gentamicin   >6        6          ≦4                                   Imipenem     >8        8          ≦4                                   Nitrofurantoin                                                                             >64       64          ≦32                                 Norfloxacin  >8        8          ≦4                                   Oxacillin    ≧4            ≦2                                   Penicillin     ≧0.25         ≦0.12                              Rifampin     >2        2          ≦1                                   Sulfamethoxazole                                                                           >256                 ≦256                                 Tetracycline ≧128                                                                             8          ≦4                                   Ticarcillin/K Clav.                                                                        ≧8            ≦4                                   Trimethoprim/                                                                               >2/38                ≧2/38                               Sulfamethoxazole                                                              Vancomycin   >16        8-16      ≦4                                   ______________________________________                                    

                                      TABLE 11X                                   __________________________________________________________________________    EFFECTS OF BPI PROTEIN PRODUCTS ± ANTIBIOTICS                                              Minimum Inhibitory Concentration of Antibiotic (μg/mL)                     With:                                                         Organism                        4               16                            Name            0   4   4   4   μg/mL                                                                          16  16  16  μg/mL                      (Microscan ®                                                                     Antibiotic                                                                             μg/mL                                                                          μg/mL                                                                          μg/mL                                                                          μg/mL                                                                          rBPI.sub.42                                                                       μg/mL                                                                          μg/mL                                                                          μg/mL                                                                          rBPI.sub.42                   ID No.)                                                                              Tested   BPI rBPI.sub.21                                                                       rBPI.sub.23                                                                       rBPI.sub.50                                                                       dimer                                                                             rBPI.sub.21                                                                       rBPI.sub.23                                                                       rBPI.sub.50                                                                       dimer                         __________________________________________________________________________    S. aureus                                                                            Cephalothin                                                                            8   4   4   4   8   4   4   4   4                             052-106                                                                              Cefuroxime**                                                                           >16 16  16  16  >16 8   8   8   8                                    Ceftriaxone*                                                                           16  8   8   8   16  8   8   8   8                                    Chloramphenicol                                                                        8   8   8   8   8   <4  <4  16  <4                                   Trimethoprim/                                                                          >2  <2  <2  <2  <2  <2  <2  <2  <2                                   Sulfamethoxazole**                                                            Ciprofloxacin                                                                          <1  <1  <1  <1  <1  <1  <1  >2  <1                                   Sulfamethoxazole**                                                                     >256                                                                              >256                                                                              >256                                                                              >256                                                                              >256                                                                              <256                                                                              >256                                                                              >256                                                                              <256                          __________________________________________________________________________

EXAMPLE 17 In Vitro Effects of BPI Protein products Alone or inCombination with Antibiotics on Strains of the Gram-Positive OrganismStaphylococcus Epidermidis

The effect of a BPI protein product, rBPI₂₁, on the antibioticsusceptibility of a variety of Staphylococcus epidermidis species wasevaluated using the Microscan® antibiotic susceptibility screening assayof Example 9. The direct growth inhibitory effect of rBPI₂₁ on thesespecies was also evaluated in the same assay. Assays were conducted onthe following clinical isolates of S. epidermidis (from BaxterMicroscan® library, Sacramento, Calif.): 055051, 055155, 19776, 20778,20959, 055125, 055129, 055126, 32086 and 32085. S. epidermidis is acoagulase-negative staphylococcus that is a major agent of nosocomial(hospital-acquired) sepsis in oncology and neonatal sections, andaccounts for about 40% of all prosthetic joint infections.

These strains of S. epidermidis were tested to determine the MIC₉₀ foreach antibiotic in the panel at varying concentrations of rBPI₂₁.Results are reported in Table 12 below, and applicable antibioticsusceptibility standards (interpretation of an MIC as clinicallyresistant, intermediate or susceptible according to NCCLS standards)appear in Table 12A.

The data show that the BPI protein product was able to reduce the S.epidermidis MIC₉₀ values for thirteen of twenty-five antibioticsevaluated: amikacin, amoxicillin/K clavulanate (augmentin), ampicillin,cefazolin, cefotaxime, ceftriaxone, cefuroxime, chloramphenicol,penicillin, sulfamethoxazole, tetracycline, ticarcillin/K clavulanateand trimethoprim/sulfamethoxazole. None of the S. epidermidis strainstested were susceptible to BPI protein product alone, withoutantibiotic.

                  TABLE 12                                                        ______________________________________                                        Effect of rBPI.sub.21 on MIC.sub.90 of Antibiotics                            for Staphylococcus epidermidis                                                           MIC.sub.90 With                                                                          MIC.sub.90 With                                                                          MIC.sub.90 With                                         0 μg/mL rBPI.sub.21                                                                   8 μg/mL rBPI.sub.21                                                                   32 μg/mL rBPI.sub.21                      Antibiotic Tested                                                                        (n = 35).sup.a                                                                           (n = 10).sup.a                                                                           (n = 9).sup.a                                ______________________________________                                        Amikacin   32         NT         16                                           Amoxicillin/K Clav                                                                       8          8          4                                            Ampicillin/                                                                              8          NT         8                                            Sulbactam                                                                     Ampicillin --         --         4                                            Cefazolin  4          4          2                                            Cefotaxime --         --         8                                            Ceftriaxone                                                                              --         NT         4                                            Cefuroxime --         NT         2                                            Cephalothin                                                                              8          8          8                                            Chloramphenicol                                                                          --         NT         4                                            Ciprofloxacin                                                                            1          1          1                                            Clindamycin                                                                              --         --         --                                           Erythromycin                                                                             --         --         --                                           Gentamicin --         --         --                                           Imipenem   --         --         --                                           Nitrofurantoin                                                                           32         32         32                                           Norfloxacin                                                                              4          4          4                                            Oxacillin  --         --         --                                           Penicillin --         --         8                                            Rifampin   1          1          1                                            Sulfamethoxazole                                                                         --         NT         256                                          Tetracycline                                                                             128        128        8                                            Ticarcillin/K Clav                                                                       --         NT         1                                            Trimeth/Sulfa                                                                            --         2          2                                            Vancomycin 2          2          2                                            ______________________________________                                         .sup.a Number of runs.                                                   

                  TABLE 12A                                                       ______________________________________                                        Susceptibility ranges for S. epidermidis                                                 MIC (μg/mL)                                                     Antibiotic   Resistant Intermediate                                                                             Susceptible                                 ______________________________________                                        Amikacin     >32       32         ≦16                                  Amoxicillin/K Clav.                                                                        ≧8/4          ≦4/2                                 Ampicillin/Sulbactam                                                                       ≦16/8                                                                            16/8       ≦8/4                                 Ampicillin   ≧0.5          ≦0.25                                Cefazolin    >16       16         ≦8                                   Cefotaxime   >32       16-32      ≦8                                   Ceftriaxone  >32       16-32      ≦8                                   Cefuroxime   >16       16         ≦8                                   Cephalothin  >16       16         ≦8                                   Chloramphenicol                                                                            >16       16         ≦8                                   Ciprofloxacin                                                                              >2         2         ≦1                                   Clindamycin  >2        1-2        ≦0.5                                 Erythromycin >4        1-4        ≦0.5                                 Gentamicin   >6         6         ≦4                                   Imipenem     >8         8         ≦4                                   Nitrofurantoin                                                                             >64       64         ≦32                                  Norfloxacin  >8         8         ≦4                                   Oxacillin    ≧4            ≦2                                   Penicillin   ≧0.25         ≦0.12                                Rifampin     >2         2         ≦1                                   Sulfamethoxazole                                                                           >256                 ≦256                                 Tetracycline ≧128                                                                              8         ≦4                                   Ticarcillin/K Clav                                                                         ≧8            ≦4                                   Trimethoprim/                                                                              >2/38                ≦2/38                                Sulfamethoxazole                                                              Vancomycin   >16        8-16      ≦4                                   ______________________________________                                    

EXAMPLE 18 In Vitro Effects of BPI Protein Products Alone or inCombination with Antibiotics on Species of the Gram-Positive OrganismStaphylococcus

The effect of a BPI protein product, rBPI₂₁, on the antibioticsusceptibility of a variety of other coagulase-negative Staphylococcusspecies was evaluated using the Microscan® antibiotic susceptibilityscreening assay of Example 9. The direct growth inhibitory effect ofrBPI₂₁ on these species was also evaluated in the same assay. Assayswere conducted on clinical isolates of Staphylococcus species (fromBaxter Microscan® library, Sacramento, Calif.). In the last decade therehas been a marked increase in clinical infections caused by thecoagulase-negative staphylococci. These organisms are also significantopportunistic pathogens.

A summary of the results of the antibiotic screening panels, reported asMICs (μg/ml) of the antibiotic tested, is shown in Table 13 below. Theantibiotic susceptibility standards (interpretation of an MIC asclinically resistant, intermediate or susceptible according to NCCLSstandards) applicable to the organism tested appear in Table 13A.

These results show that BPI protein product reversed resistance topenicillin for one S. hominis strain, and reversed resistance totrimethoprim/sulfamethoxazole for one S. haemolyticus strain and the S.intermedius strain. BPI protein product increased susceptibility of S.hominis strains to ampicillin and penicillin, the S. sciuri strain toclindamycin and penicillin, the S. saprophyticus strain to erythromycin,S. haemolyticus strains to erythromycin, and the S. hyicus strain toclindamycin and erythromycin, the S. intermedius strain to erythromycin,and the S. simulans strain to erythromycin. None of these tested strainswere susceptible to BPI₂₁ alone at the concentrations tested.

                  TABLE 13                                                        ______________________________________                                        Effects of rBPI.sub.21 on Staphylococcus Species                                               Minimum Inhibitory                                                            Concentration (μg/ml)                                                            With 0  With 4 With 16                                 Microscan  Antibiotic  μg/ml                                                                              μg/ml                                                                             μg/ml                                Library ID No..sup.a                                                                     Tested.sup.b                                                                              rBPI.sub.21                                                                           rBPI.sub.21                                                                          rBPI.sub.21                             ______________________________________                                        ( S. hominis)                                                                            Ampicillin  0.25    <0.12  <0.12                                   057-003    Penicillin**                                                                              0.5     0.06   0.06                                    (S. hominis)                                                                             Ampicillin  0.25    0.25   <0.12                                   057-001    Penicillin  0.25    0.12   0.06                                    (S. hominis)                                                                             Ampicillin  0.25    0.5    0.25                                    057-024    Penicillin  0.5     0.5    0.5                                     (S. scirui)                                                                              Pencillin   0.06    0.06   <0.03                                   060-016    Clindamycin*                                                                              1       1      0.5                                     (S. saprophyticus)                                                                       Erythromycin                                                                              0.5     0.5    <0.25                                   059-003                                                                       (S. haemolyticus)                                                                        Trim/Sulfa**                                                                              >2      <2     <2                                      19770      Erythromycin                                                                              >4      4      4                                       (S. haemolyticus)                                                                        Ampicillin  0.5     1      0.5                                     056-042    Penicillin  0.25    0.5    0.25                                    (S. hyicus)                                                                              Erythromycin                                                                              0.5     <0.25  <0.25                                   10377      Clindamycin 0.5     <0.25  <0.25                                   (S. intermedius)                                                                         Trim/Sulfa**                                                                              >2      <2     <2                                      10254      Erythromycin                                                                              0.5     <0.25  <0.25                                   (S. simulans)                                                                            Erythromycin                                                                              0.5     0.5    <0.25                                   061-001                                                                       ______________________________________                                         .sup.a Two strains of S. lugdunensis (Microscan library ID Nos. 19782 and     11130) were also tested but were susceptible to all antibiotics tested.       .sup.b Strains were susceptible to all other antibiotics that were tested     but are not shown here.                                                  

                  TABLE 13A                                                       ______________________________________                                        Susceptibility ranges for Staphylococcus species                                        MIC (μg/ml)                                                      Antibiotic  Resistant  Intermediate                                                                            Susceptible                                  ______________________________________                                        Ampicillin  #0.5                 ≦0.25                                 Clindamycm; >2         1-2       ≦0.5                                  Erythromycin                                                                              >4         1-4       ≦0.5                                  Penicillin  ≧0.25         ≦0.12                                 Trimethoprim/                                                                             >2/38                ≦2/38                                 Sulfamethoxazole                                                              ______________________________________                                    

EXAMPLE 19 Early In Vitro Bactericidal Effects of BPI Protein ProductAlone on in Combination with Antibiotics on S. aureus, S. Pneumoniae,and E. faecium

The effect of a BPI protein product, rBPI₂₁, on the killing curves ofselected antibiotics was determined for selected organisms. Microscan®panel plates were prepared for a methicillin-resistant S. aureus(Microscan library ID no. 052-106), S. pneumoniae (Microscan library IDno. 31573), and E. faecium (Microscan library ID no. 15773), accordingto Example 9. Cell suspensions were added to 25 ml Pluronic InoculumWater containing 0 or 16 μg/ml rBPI₂₁. After inoculation, the panelplates were incubated at 35° C. for 24 hours. At 0, 4, 7 and 24 hoursafter inoculation, 5 μl samples were removed from each growth controlwell (containing culture media without antibiotic) and from each wellcontaining: 8 μg/ml penicillin, 2 μg/ml ciprofloxacin, 256 μg/mlsulfamethoxazole, 32 μg/ml cefotaxime, 16 μg/ml chloramphenicol, or 16μg/ml vancomycin. These 5 μl samples were diluted in sterile water andinoculated onto Trypticase Soy agar plates (Remel, Lenexa, Kans.) orblood agar plates (Remel, Lenexa, Kans.) for S. pneumoniae. After 48hours of incubation at 35° C., the plates were counted and the number ofcolony forming units of bacteria remaining in the well was calculated.

The results are shown below in FIGS. 26 through 32. In all of thefigures, the growth in the presence of antibiotic alone (without rBPI₂₁)is indicated for: S. aureus (a filled square), S. pneumoniae (a filleddiamond) or E. faecium (a filled triangle). Also, in all figures, thegrowth in the presence of antibiotic with rBPI₂₁ is indicated for: S.aureus (an open square), S. pneumoniae (an open diamond) or E. faecium(an open triangle).

FIG. 26 shows the growth curve of organisms with rBPI₂₁ (and withoutantibiotic) and without rBPI₂₁ (and without antibiotic). In FIG. 26, thegrowth curves for S. aureus with rBPI₂₁ (open squares) and E. faeciumwithout rBPI₂₁ (filled triangles) overlap at 7 and 24 hours. The resultsshow that BPI protein product has a dramatic bactericidal effect on S.pneumoniae that continues through 24 hours, and a moderate earlyinhibitory effect on growth of E. faecium that is not sustained after 10hours.

In FIG. 27, the growth curves for S. pneumoniae with rBPI₂₁ (opendiamonds) and S. pneumoniae without rBPI₂₁ (filled diamonds) overlap at7 and 24 hours. FIG. 27 shows that the BPI protein product enhanced theearly bactericidal effect of penicillin on all three organisms at 0-10hours.

In FIG. 28, the growth curves for S. pneumoniae with rBPI₂₁ (opendiamonds) and S. pneumoniae without rBPI₂₁ (filled diamonds) overlapcompletely. FIG. 28 shows that BPI protein product enhanced the earlybactericidal activity of cefotaxime for S. aureus and E. faecium at 0-10hours.

FIG. 29 shows that BPI protein product enhanced the early bactericidalactivity of chloramphenicol for all three organisms at 0-10 hours.

FIG. 30 shows that BPI protein product enhanced the early bactericidalactivity of sulfamethoxazole for all three organisms at 0-10 hours.

In FIG. 31, the growth curves for S. aureus with rBPI₂₁ (open squares),S. aureus without rBPI₂₁ (filled squares) and S. pneumoniae withoutrBPI₂₁ (filled diamonds) overlap almost completely. FIG. 31 shows thatBPI protein product enhanced the early bactericidal activity ofciprofloxacin for S. pneumoniae and E. faecium at 0-10 hours.

In FIG. 32, the growth curves for S. pneumoniae with rBPI₂₁ (opendiamonds) and S. pneumoniae without rBPI₂₁ (filled diamonds) overlapcompletely. FIG. 32 shows that BPI protein product enhanced the earlybactericidal effect of vancomycin for S. aureus and E. faecium at 0-10hours.

The early time course of the bactericidal activity of BPI proteinproduct was studied for three clinical isolates (methicillin-resistantS. aureus, S. pneumoniae, and E. faecium). Approximately 4 to 10colonies of each bacterial species, from 18 to 24 hour growth on eithertrypticase soy agar plates (for S. aureus and E. faecium) or 5% sheepblood agar plates (for S. pneumoniae) (Remel, Lenexa, Kans.), wereemulsified in sterile water to a density equivalent to an 0.5 McFarlandstandard; 100 μL of this bacterial inoculum was transferred to glasstubes containing 25 mL Microscan® Pluronic Inoculum Water, such that thefinal concentration of organisms was ˜4 to 7×10⁵ cells/mL, and rBPI₂₁was added to a final concentration of 16 μg/mL. The tubes were mixed byinversion, and a control and BPI sample were immediately removed anddiluted 1:100 in sterile water for colony counts. Similar samples weretaken at 7.5, 15, 30, 60, 90 and 120 minutes. The plates were incubatedat 35° C. to 37° C. for 15 to 24 hours and the CFUs/mL were measured bydirect counts.

Some bactericidal activity was observed for all isolates afer 15 minutesof incubation at room temperature. However, the strain consideredsusceptible to the direct bactericidal effects of BPI protein product(S. pneumoniae) was immediately reduced to 0 CFU at 7.5 minutes, whilethe killing curves for "resistant" strains (S. aureus and E. faecium)were prolonged. There was at least a three-log reduction in the numberof organisms after the BPI treatment; however, 0 CFU was never reached.CFU data is summarized in Table 14 below.

                                      TABLE 14                                    __________________________________________________________________________    Calculated CFUs/mL                                                                    0 mins                                                                            7.5 mins                                                                           15 mins                                                                           30 mins                                                                           60 mins                                                                           90 mins                                                                           120 mins                                     __________________________________________________________________________    S. pneumoniae                                                                         179000                                                                            173000                                                                             153000                                                                            187000                                                                            169000                                                                            168000                                                                            143000                                       control                                                                       S. pneumoniae                                                                         6000                                                                              2    0   0   0   686 0                                            with rBPI.sub.21                                                              S. aureus control                                                                     488000                                                                            615000                                                                             440000                                                                            640000                                                                            480000                                                                            720000                                                                            410000                                       S. aureus with                                                                        650000                                                                            5000 300 260 510 530 510                                          rBPI.sub.21                                                                   E. faecium                                                                            276000                                                                            193000                                                                             265000                                                                            277000                                                                            265000                                                                            301000                                                                            212000                                       control                                                                       E. faecium with                                                                       131000                                                                            129000                                                                             41000                                                                             3340                                                                              990 700 570                                          rBPI.sub.21                                                                   __________________________________________________________________________

To determine if the reason S. aureus organisms had survived the initialBPI treatment was due to resistance to BPI protein product, coloniesrecovered after 90 minutes of incubation with 16 μg/mL rBPI₂₁ were grownfor 24 hours, prepared as previously described and incubated with anadditional 16 μg/mL rBPI₂₁. Control S. aureus that had not beenpreviously exposed to BPI protein product were also incubated with 16μg/mL rBPI₂₁. After 30 minutes of incubation, the survivors and controlswere plated and counted. As seen in Table 15 below, the survivors froman initial treatment with rBPI₂₁ were not resistant to BPI proteinproduct.

                  TABLE 15                                                        ______________________________________                                                      Control    16 μg/ml rBPI.sub.21                              ______________________________________                                        S. aureus       5 × 10.sup.5 CFU/mL                                                                  3 × 10.sup.2 CFU/mL                        S. aureus recovered after                                                                     4 × 10.sup.5 CFU/mL                                                                  2 × 10.sup.2 CFU/mL                        90 min. incubation with                                                       16 μg/mL rBPI.sub.21                                                       ______________________________________                                    

In another experiment, S. aureus organisms were incubated for 30 minutesin 16 μg/mL rBPI₂₁, and an aliquot was removed, plated and counted. Halfof the BPI-treated cell suspension was incubated with an additional 16μg/mL rBPI₂₁. Incubation continued for both cell suspensions for anadditional 30 minutes. The additional rBPI2, treatment decreased therelative number of survivors over the untreated control (84%reduction/30 min. vs 34% control reduction/30 min.).

It should be noted that incubation in Mueller-Hinton broth inhibited thecomplete killing of S. pneumoniae between 0 and 7.5 minutes. Inadditional experiments conducted to explore this phenomenon, it wasdetermined that the addition of calcium chloride to the incubationmedium appeared to reduce the effectiveness of rBPI₂₁ as measured byincreased CFUs. A preliminary experiment indicating that NaCl may alsoreduce the effectiveness of rBPI₂₁ (i.e., increased CFUs) suggests thatosmolality may be a factor.

EXAMPLE 20 In Vitro Effect of BPI Protein Products Alone or inCombination with Antibiotics on Gram-Positive Organisms

BPI protein products are evaluated, using a multiwell antibioticsusceptibility screening assay, for their effects alone or incombination with penicillin, ampicillin or ciprofloxacin on a culture ofStreptococcus pneumoniae (especially penicillin-resistant organisms).BPI protein products are also evaluated alone or in combination with theantibiotics vancomycin, rifampin, ciprofloxacin, cefazolin,vancomycin/gentamicin and ciprofloxacin/pip eracillin for their effectson the organism Staphylococcus aureus (especially methicillin resistantorganisms). BPI protein products are evaluated alone or in combinationwith penicillin, ampicillin, vancomycin, ciprofloxacin,penicillin/gentamicin, or azithromycin for their effects on Enterococcus(multiply resistant strains). The effect of BPI protein products aloneor in combination with vancomycin or clindamycin on Corynebacteria areevaluated.

EXAMPLE 21 In Vitro Enhancement of the Bactericidal Activity of BPIProtein Products by Selected Poloxamers

The anti-bacterial activity of therapeutic compositions comprising a BPIprotein product and a variety of different poloxamer surfactants wasevaluated as described in co-owned, co-pending U.S. patent applicationSer. No. 08/754,998, filed concurrently herewith (Attorney Docket No.27129/32424), now abandoned, the disclosure of which is hereinincorporated by reference. Briefly, therapeutic compositions comprisinga BPI protein product and a poloxamer surfactant at concentrationsranging from 0.005 to 0.1% (weight/volume) were prepared and incubatedat 37° C. with gram-positive organisms in water, broth, or varyingconcentrations of serum. After incubation, the colony forming units(CFU) of bacteria remaining were determined to ascertain enhancement ofanti-bacterial activity. Studies were also performed using poloxamersurfactants that had been heat-treated using the following procedure:(1) making a solution of the poloxamer in deionized water, (2) heatingthe solution to a boil, (3) removing it from heat, (4) allowing it tocool to room temperature, and (5) stirring until the poloxamer iscompletely solubilized. Alternatively, in the heating step (2), thesolution may be boiled for up to 30 minutes or more.

It was observed that poloxamer 333 (heat-treated or not) and poloxamer403 (heat-treated or not) and poloxamer 335 and heat-treated poloxamer334 provided enhancement of the anti-bacterial activity of BPI proteinproducts against bacteria.

EXAMPLE 22 In Vivo Effects of BPI Protein Products Alone or withAntibiotics

Preliminary experiments, in which BPI protein products alone or withpenicillin were administered to mice challenged intravenously with 5×10⁸CFU of the S. aureus bacterial form of Example 3 above, did not show aneffect on mortality. This outcome was not unexpected in view of the lackof effect of BPI protein products on that bacterial form in the in vitroassays of Examples 2, 3 and 16. No effect on mortality was observed inpreliminary experiments in which mice were challenged with 1×10⁵ CFU ofa S. pneumoniae bacterial form which appeared in a preliminaryexperiment to be susceptible to rBPI₂₁ alone in the Microscan® system.Additional in vivo experiments will be performed in a variety of otheranimal models, including using a variety of different BPI proteinproducts alone and in conjunction with a variety of differentantibiotics.

A. EVALUATION IN THE MOUSE PNEUMONIA MODEL

The effect on gram-positive infections of BPI protein productsadministered alone or in combination with antibiotics is evaluated in amouse pneumonia model. Pneumonia is induced in mice by intranasalinstillation of 3 drops of an undiluted overnight culture of S.pneumoniae in trypticase soy broth containing 5% goat serum. Thechallenge dose is administered under light Metofane anesthesia with a1-ml tuberculin syringe and a 22-gauge needle. Infected mice are treated24 hours after infection with a single intravenous dose of BPI proteinproduct alone, antibiotic alone, or the combination of both agents. Themice are sacrificed 48 hours after infection. The lungs are removedaseptically and cultured for the presence of organism by touching a cutsurface of the lung to a blood agar plate. Under these conditions, at 48hours after infection, untreated controls exhibit consolidation of thelungs and heavily positive cultures. Further, in this model, most of theuntreated controls die within 96 hours after infection.

B. EVALUATION IN THE RABBIT ENDOCARDITIS MODEL

The effect on gram-positive infections of BPI protein productsadministered alone or in combination with antibiotics is evaluated in arabbit endocarditis model. Young New Zealand white rabbits weighing 1 to2 kg are anesthetized by the intravenous injection of 40 to 60 mg ofpentobarbitone. An incision is made parallel to the trachea,approximately 4 cm long and 1 cm to the right of the midline. To produceright-sided endocarditis, the jugular vein is exposed and opened betweenligatures. The lower ligature is loosened, and a polyethylene catheterof 0.8 mm external diameter and 0.4 mm internal diameter containingsterile saline is passed toward the heart until resistance and pulsationindicates that it has entered the right atrium or ventricle. Thecatheter is then secured in place by tightening both ligatures, anyexcess is cut off, and the distal end is sealed with a heated spatulabefore the skin is closed over the catheter with silk sutures Forleft-sided endocarditis, the right carotid artery can be exposed througha similar incision, opened between ligatures, and a catheter passedtoward the heart until pulsation resistance and reflux of arterial bloodindicates that it has reached the aortic valve or passed beyond into theleft ventricle. It is then secured as described above.

The presence of a catheter in the heart results in the development ofsterile vegetations consisting of small, rough whitish nodules 1 to 2 mmin size, usually at points of contact between the catheter and theendocardium. The sterile vegetations are infected by a single injectionof bacteria into an ear vein. If Streptococcus viridans is used forinfection, the infecting inoculum can be prepared by diluting anovernight glucose broth culture and injecting approximately 10⁸organisms in a volume of 1 ml. Other gram-positive organisms can be usedand may include Micrococcus albus, S. aureus, S. epidermidis, and otherstrains. An infecting inoculum can be prepared for these organisms bypreparing a 1:10 dilution of an overnight glucose broth culture andinjecting 1 mL intravenously. BPI protein product alone, antibioticalone, or the combination treatment is given intravenously in dailydoses. After days, the rabbits are sacrificed and any vegetations foundare removed aseptically. The vegetations are weighed and homogenized inglucose broth. Bacterial counts are determined by preparing serialdilutions in glucose broth and incorporating the dilutions into bloodagar pour plates. The CFU of organism per g of wet weight is thencalculated.

C. EVALUATION IN THE MOUSE INTRAPERITONEAL ABSCESS MODEL

The effect on gram-positive infections of BPI protein productsadministered alone or in combination with antibiotics is evaluated in amouse intraperitoneal abscess model. S. aureus is grown for 24 hours intrypticase soy broth with constant shaking. Cultures are continuouslygassed with 100% oxygen to minimize α-hemolysin production. Theorganisms are harvested by centrifugation, washed in saline contaning 1%trypticase soy broth (v/v), and resuspended in the same diluent toapproximately 1×10¹⁰ to 5×10¹⁰ bacteria per ml. Bacterial counts can beconfirmed by plate counts.

Groups of female white Swiss mice weighing 20 to 30 g are inoculatedintraperitoneally with 0.5 ml of a suspension contaning 1 to 2×10⁹bacteria. Beginning at 3 hours and at various intervals thereafter,subgroups of infected animals selected at random are sacrificed, and theclumped organisms or abscesses are aseptically removed from theperitoneal cavities. All clumps or abscesses from a single animal areground and homogenized in 5 ml of saline containing 1% trypticase soybroth. The suspension is serially diluted and the bacterial populationdetermined by plate counts.

D. EVALUATION IN THE MOUSE THIGH ABSCESS MODEL

The effect on gram-positive infections of BPI protein productsadministered alone or in combination with antibiotics is evaluated in amouse thigh abscess model.

The thigh lesion model provides a nonlethal experimental infection toevaluate the effectiveness of an antimicrobial and allows themeasurement of drug-pathogen interaction and drug pharmacokinetics inthe infected host. If the animals are made neutropenic, then the thighmodel becomes an excellent system for measuring the drug-microorganisminteraction with most of the host defense system eliminated.

Swiss outbred mice, preferably ICR mice, weighing 23 to 27 g are used.The mice are made neutropenic by administering cyclophosphamide (150 and100 mg/kg i.p.) on days 0 and 3, respectively. By day 4 severeneutropenia is induced (<100 neutrophils/mm³) which lasts for two tothree days. The infecting organism is grown in broth to log phase andadjusted to an OD of 0.30 at 580 nm (˜10⁶ to 10⁷ CFU/ml). The mice areinfected on day 4 by injecting 0.1 ml of this inoculum into each thighwhile the animals are under light ether anesthesia. The infection isallowed to proceed for 2 hours. The antimicrobial is then administeredin graded doses subcutaneously to groups of infected mice. Infected,untreated animals serve as controls. Two to four mice from eachtreatment group are sacrificed every hour for the first 4 hours andevery 2 to 4 hours until 16 hours after treatment. Infected, untreatedanimals are killed at similar times.

The thigh muscles are removed at each sampling period and homogenizedimmediately in 9 ml of 0.85% NaCl with a Polytron tissue homogenizer.Viable counts are determined after plating duplicate 10μ samples ofserial 10-fold dilutions of the homogenates on appropriate media. Thelog CFU/thigh is determined at each time point for groups of treated anduntreated animals.

E. EVALUATION IN A RABBIT INTRAOCULAR INFECTION MODEL

The effect on gram-positive infections of BPI protein productsadministered alone or in combination with antibiotics is evaluated inthe following model. The rabbits' eyes, anesthetized locally with 0.5%tetracaine hydrochloride, are sterilized with Metaphen (1:4000) andflushed with normal saline. Using a 26-gauge needle, approximately 0.2ml containing 300,000 or 5,000 CFU or 0.02 ml containing 700 CFU of S.aureus is inoculated into the center of the rabbit cornea, the anteriorchamber, or the vitreous of the eye. Cultures are taken at 24, 48, and72 hours from the corneas and anterior chambers of the rabbits' eyesthat received corneal inoculations. All corneas are flushed withMetaphen (1:4000) and saline before culturing to eliminate surfacecontaminants.

Cultures are taken at the same intervals from the anterior chamber andvitreous of rabbit eyes that were inoculated in the anterior chamber.Vitreous humor is removed with a 19-gauge needle. The irises are alsocultured.

The eyes that are inoculated intravitreally are also cultured at 24, 48,and 72 hours. Samples of the vitreous humor and anterior chamber as wellas the retina are used to determine the progress of the infection.

When 300,000 CFU in 0.2 ml of broth is inoculated into the corneas,anterior chamber, and vitreous of the rabbit's eyes, a virulentpanophthalmitis is generally produced within 24 to 48 hours, anddestruction of the eye occurs within 72 hours regardless of the site ofinoculation. When 5000 CFU in 0.2 ml is inoculated into the corneas,anterior chambers, and vitreous, a panophthalmitis generally results in72 hours, with infections being most severe following intravitrealinoculations and less intense when the anterior chamber is the site ofinoculation. When 700 CFU in 0.02 ml is used as the inoculum, theinfections are generally eliminated in the corneas and anterior chamberswithin 24 hours but not in the vitreous.

Other models of ocular infection and disease known in the art see, e.g.,Sugar et al., Arch. Opthalmol., 104:1230-1232 (1986) and Moon et al.,Investigative Ophthalmol. Visual Science, 29:1277-1284 (1988)! may beused for the assessment of BPI protein product effects on ocularconditions associated with bacterial infection, when the BPI proteinproduct is administered alone or in conjunction with antibiotics.

Numerous modifications and variations in the practice of the inventionare expected to occur to those skilled in the art upon consideration ofthe foregoing description of the presently preferred embodimentsthereof. Consequently, the only limitations which should be placed uponthe scope of the present invention are those which appear in theappended claims.

    __________________________________________________________________________    SEQUENCE LISTING                                                              (1) GENERAL INFORMATION:                                                      (iii) NUMBER OF SEQUENCES: 237                                                (2) INFORMATION FOR SEQ ID NO:1:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 29 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "Domain I"                                             (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                                       AlaSerGlnGlnGlyThrAlaAlaLeuGlnLysGluLeuLysArgIle                              151015                                                                        LysIleProAspTyrSerAspSerPheLysIleLysHis                                       2025                                                                          (2) INFORMATION FOR SEQ ID NO:2:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 30 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.14"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                                       GlyThrAlaAlaLeuGlnLysGluLeuLysArgIleLysIleProAsp                              151015                                                                        TyrSerAspSerPheLysIleLysHisLeuGlyLysGlyHis                                    202530                                                                        (2) INFORMATION FOR SEQ ID NO:3:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 22 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.4"                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                                       LeuGlnLysGluLeuLysArgIleLysIleProAspTyrSerAspSer                              151015                                                                        PheLysIleLysHisLeu                                                            20                                                                            (2) INFORMATION FOR SEQ ID NO:4:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.1"                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                                       GlnGlnGlyThrAlaAlaLeuGlnLysGluLeuLysArgIleLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:5:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.54"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                                       GlyThrAlaAlaLeuGlnLysGluLeuLysArgIleLysIlePro                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:6:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 35 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "Domain II"                                            (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                                       SerSerGlnIleSerMetValProAsnValGlyLeuLysPheSerIle                              151015                                                                        SerAsnAlaAsnIleLysIleSerGlyLysTrpLysAlaGlnLysArg                              202530                                                                        PheLeuLys                                                                     35                                                                            (2) INFORMATION FOR SEQ ID NO:7:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.2"                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                                       IleLysIleSerGlyLysTrpLysAlaGlnLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:8:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.8"                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                                       LysTrpLysAlaGlnLysArgPheLeuLys                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:9:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 16 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.58"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                                       CysIleLysIleSerGlyLysTrpLysAlaGlnLysArgPheLeuLys                              151015                                                                        (2) INFORMATION FOR SEQ ID NO:10:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 17 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.65 oxidized"                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                                      CysIleLysIleSerGlyLysTrpLysAlaGlnLysArgPheLeuLys                              151015                                                                        Cys                                                                           (2) INFORMATION FOR SEQ ID NO:11:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 27 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.3"                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                                      AsnValGlyLeuLysPheSerIleSerAsnAlaAsnIleLysIleSer                              151015                                                                        GlyLysTrpLysAlaGlnLysArgPheLeuLys                                             2025                                                                          (2) INFORMATION FOR SEQ ID NO:12:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 28 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "Domain III"                                           (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                                      ValHisValHisIleSerLysSerLysValGlyTrpLeuIleGlnLeu                              151015                                                                        PheHisLysLysIleGluSerAlaLeuArgAsnLys                                          2025                                                                          (2) INFORMATION FOR SEQ ID NO:13:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 13 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.11"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                                      LysSerLysValTrpLeuIleGlnLeuPheHisLysLys                                       1510                                                                          (2) INFORMATION FOR SEQ ID NO:14:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 29 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.12"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                                      SerValHisValHisIleSerLysSerLysValGlyTrpLeuIleGln                              151015                                                                        LeuPheHisLysLysIleGluSerAlaLeuArgAsnLys                                       2025                                                                          (2) INFORMATION FOR SEQ ID NO:15:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.13"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                                      LysSerLysValGlyTrpLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:16:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.15"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                                      AlaLysIleSerGlyLysTrpLysAlaGlnLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:17:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.16"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                                      IleAlaIleSerGlyLysTrpLysAlaGlnLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:18:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.17"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                                      IleLysAlaSerGlyLysTrpLysAlaGlnLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:19:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.18"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                                      IleLysIleAlaGlyLysTrpLysAlaGlnLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:20:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.19"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                                      IleLysIleSerAlaLysTrpLysAlaGlnLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:21:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.20"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                                      IleLysIleSerGlyAlaTrpLysAlaGlnLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:22:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.21"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                                      IleLysIleSerGlyLysAlaLysAlaGlnLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:23:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.22"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:                                      IleLysIleSerGlyLysTrpAlaAlaGlnLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:24:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.23"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:                                      IleLysIleSerGlyLysTrpLysAlaAlaLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:25:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.24"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:                                      IleLysIleSerGlyLysTrpLysAlaGlnAlaArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:26:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.25"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:                                      IleLysIleSerGlyLysTrpLysAlaGlnLysAlaPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:27:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.26"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:                                      IleLysIleSerGlyLysTrpLysAlaGlnLysArgAlaLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:28:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.27"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:                                      IleLysIleSerGlyLysTrpLysAlaGlnLysArgPheAlaLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:29:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.28"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:                                      IleLysIleSerGlyLysTrpLysAlaGlnLysArgPheLeuAla                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:30:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.59"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:                                      IleLysIleSerGlyAlaTrpAlaAlaGlnLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:31:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.45"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:                                      IleLysIleSerGlyLysTrpLysAlaAlaAlaArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:32:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.60"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:                                      IleAlaIleSerGlyLysTrpLysAlaGlnLysArgPheLeuAla                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:33:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.31"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:                                      AlaSerLysValGlyTrpLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:34:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.32"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:                                      LysAlaLysValGlyTrpLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:35:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.33"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:                                      LysSerAlaValGlyTrpLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:36:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.34"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:                                      LysSerLysAlaGlyTrpLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:37:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.35"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:                                      LysSerLysValAlaTrpLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:38:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.36"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:                                      LysSerLysValGlyAlaLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:39:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.37"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:                                      LysSerLysValGlyTrpAlaIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:40:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.38"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:                                      LysSerLysValGlyTrpLeuAlaGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:41:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.39"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:                                      LysSerLysValGlyTrpLeuIleAlaLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:42:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.40"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:                                      LysSerLysValGlyTrpLeuIleGlnAlaPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:43:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.41"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43:                                      LysSerLysValGlyTrpLeuIleGlnLeuAlaHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:44:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.42"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:                                      LysSerLysValGlyTrpLeuIleGlnLeuPheAlaLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:45:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.43"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45:                                      LysSerLysValGlyTrpLeuIleGlnLeuPheHisAlaLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:46:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.44"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46:                                      LysSerLysValGlyTrpLeuIleGlnLeuPheHisLysAla                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:47:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.56"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:47:                                      IleLysIleSerGlyLysTrpLysAlaLysGlnArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:48:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.61"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48:                                      IleLysIleSerGlyLysPheLysAlaGlnLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:49:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.66"                                               (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 7                                                               (D) OTHER INFORMATION: /label=D- Trp                                          /note="The amino acid at position 7 is                                        D- tryptophan"                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49:                                      IleLysIleSerGlyLysTrpLysAlaGlnLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:50:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.67"                                               (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6..8                                                            (D) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 7 is                                           beta-1- naphthyl-substituted"                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:                                      IleLysIleSerGlyLysAlaLysAlaGlnLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:51:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.9"                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:51:                                      LysArgPheLeuLysLysTrpLysAlaGlnLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:52:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.30"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52:                                      LysTrpLysAlaGlnLysArgPheLeuLysLysSerLysValGlyTrp                              151015                                                                        LeuIleGlnLeuPheHisLysLys                                                      20                                                                            (2) INFORMATION FOR SEQ ID NO:53:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 29 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.63"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53:                                      IleLysIleSerGlyLysTrpLysAlaGlnLysArgPheLeuLysLys                              151015                                                                        SerLysValGlyTrpLeuIleGlnLeuPheHisLysLys                                       2025                                                                          (2) INFORMATION FOR SEQ ID NO:54:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.7"                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54:                                      LysTrpLysAlaGlnLysArgPheLeuLysLysTrpLysAlaGlnLys                              151015                                                                        ArgPheLeuLys                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:55:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 25 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.10.1"                                             (xi) SEQUENCE DESCRIPTION: SEQ ID NO:55:                                      LysArgPheLeuLysLysTrpLysAlaGlnLysArgPheLeuLysLys                              151015                                                                        TrpLysAlaGlnLysArgPheLeuLys                                                   2025                                                                          (2) INFORMATION FOR SEQ ID NO:56:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 28 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.29"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:56:                                      LysSerLysValGlyTrpLeuIleGlnLeuPheHisLysLysLysSer                              151015                                                                        LysValGlyTrpLeuIleGlnLeuPheHisLysLys                                          2025                                                                          (2) INFORMATION FOR SEQ ID NO:57:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.46"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57:                                      LysTrpLysAlaAlaAlaArgPheLeuLysLysTrpLysAlaGlnLys                              151015                                                                        ArgPheLeuLys                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:58:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.47"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58:                                      LysTrpLysAlaGlnLysArgPheLeuLysLysTrpLysAlaAlaAla                              151015                                                                        ArgPheLeuLys                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:59:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.48"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59:                                      LysTrpLysAlaAlaAlaArgPheLeuLysLysTrpLysAlaAlaAla                              151015                                                                        ArgPheLeuLys                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:60:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 30 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.69"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:60:                                      LysTrpLysAlaAlaAlaArgPheLeuLysLysTrpLysAlaAlaAla                              151015                                                                        ArgPheLeuLysLysTrpLysAlaAlaAlaArgPheLeuLys                                    202530                                                                        (2) INFORMATION FOR SEQ ID NO:61:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 21 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.55"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:61:                                      GlyTrpLeuIleGlnLeuPheHisLysLysIleGluSerAlaLeuArg                              151015                                                                        AsnLysMetAsnSer                                                               20                                                                            (2) INFORMATION FOR SEQ ID NO:62:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.73"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62:                                      IleLysIleSerGlyLysTrpLysAlaGlnPheArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:63:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.70"                                               (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 8..10                                                           (D) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 7 is                                           beta-3- pyridyl-substituted"                                                  (xi) SEQUENCE DESCRIPTION: SEQ ID NO:63:                                      IleLysIleSerGlyLysAlaLysAlaGlnLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:64:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.71"                                               (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 13..15                                                          (D) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 13 is                                          beta-3- pyridyl-substituted"                                                  (xi) SEQUENCE DESCRIPTION: SEQ ID NO:64:                                      IleLysIleSerGlyLysTrpLysAlaGlnLysArgAlaLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:65:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 26 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.10.2"                                             (xi) SEQUENCE DESCRIPTION: SEQ ID NO:65:                                      GlnLysArgPheLeuLysLysTrpLysAlaGlnLysArgPheLeuLys                              151015                                                                        LysTrpLysAlaGlnLysArgPheLeuLys                                                2025                                                                          (2) INFORMATION FOR SEQ ID NO:66:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 17 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.72"                                               (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 1..3                                                            (D) OTHER INFORMATION: /label=D- alanine                                      /note="The position 1 and position 2 alanine                                  residues are both D-alanine"                                                  (xi) SEQUENCE DESCRIPTION: SEQ ID NO:66:                                      AlaAlaIleLysIleSerGlyLysTrpLysAlaGlnLysArgPheLeu                              151015                                                                        Lys                                                                           (2) INFORMATION FOR SEQ ID NO:67:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 22 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.5"                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:67:                                      ValHisValHisIleSerLysSerLysValGlyTrpLeuIleGlnLeu                              151015                                                                        PheHisLysLysIleGlu                                                            20                                                                            (2) INFORMATION FOR SEQ ID NO:68:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 17 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.65 reduced"                                       (ix) FEATURE:                                                                 (A) NAME/KEY: Disulfide-bond                                                  (B) LOCATION: 1..17                                                           (xi) SEQUENCE DESCRIPTION: SEQ ID NO:68:                                      CysIleLysIleSerGlyLysTrpLysAlaGlnLysArgPheLeuLys                              151015                                                                        Cys                                                                           (2) INFORMATION FOR SEQ ID NO:69:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 487 amino acids                                                   (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "rBPI"                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:69:                                      MetArgGluAsnMetAlaArgGlyProCysAsnAlaProArgTrpVal                              31-30-25-20                                                                   SerLeuMetValLeuValAlaIleGlyThrAlaValThrAlaAlaVal                              15-10-51                                                                      AsnProGlyValValValArgIleSerGlnLysGlyLeuAspTyrAla                              51015                                                                         SerGlnGlnGlyThrAlaAlaLeuGlnLysGluLeuLysArgIleLys                              202530                                                                        IleProAspTyrSerAspSerPheLysIleLysHisLeuGlyLysGly                              354045                                                                        HisTyrSerPheTyrSerMetAspIleArgGluPheGlnLeuProSer                              50556065                                                                      SerGlnIleSerMetValProAsnValGlyLeuLysPheSerIleSer                              707580                                                                        AsnAlaAsnIleLysIleSerGlyLysTrpLysAlaGlnLysArgPhe                              859095                                                                        LeuLysMetSerGlyAsnPheAspLeuSerIleGluGlyMetSerIle                              100105110                                                                     SerAlaAspLeuLysLeuGlySerAsnProThrSerGlyLysProThr                              115120125                                                                     IleThrCysSerSerCysSerSerHisIleAsnSerValHisValHis                              130135140145                                                                  IleSerLysSerLysValGlyTrpLeuIleGlnLeuPheHisLysLys                              150155160                                                                     IleGluSerAlaLeuArgAsnLysMetAsnSerGlnValCysGluLys                              165170175                                                                     ValThrAsnSerValSerSerLysLeuGlnProTyrPheGlnThrLeu                              180185190                                                                     ProValMetThrLysIleAspSerValAlaGlyIleAsnTyrGlyLeu                              195200205                                                                     ValAlaProProAlaThrThrAlaGluThrLeuAspValGlnMetLys                              210215220225                                                                  GlyGluPheTyrSerGluAsnHisHisAsnProProProPheAlaPro                              230235240                                                                     ProValMetGluPheProAlaAlaHisAspArgMetValTyrLeuGly                              245250255                                                                     LeuSerAspTyrPhePheAsnThrAlaGlyLeuValTyrGlnGluAla                              260265270                                                                     GlyValLeuLysMetThrLeuArgAspAspMetIleProLysGluSer                              275280285                                                                     LysPheArgLeuThrThrLysPhePheGlyThrPheLeuProGluVal                              290295300305                                                                  AlaLysLysPheProAsnMetLysIleGlnIleHisValSerAlaSer                              310315320                                                                     ThrProProHisLeuSerValGlnProThrGlyLeuThrPheTyrPro                              325330335                                                                     AlaValAspValGlnAlaPheAlaValLeuProAsnSerSerLeuAla                              340345350                                                                     SerLeuPheLeuIleGlyMetHisThrThrGlySerMetGluValSer                              355360365                                                                     AlaGluSerAsnArgLeuValGlyGluLeuLysLeuAspArgLeuLeu                              370375380385                                                                  LeuGluLeuLysHisSerAsnIleGlyProPheProValGluLeuLeu                              390395400                                                                     GlnAspIleMetAsnTyrIleValProIleLeuValLeuProArgVal                              405410415                                                                     AsnGluLysLeuGlnLysGlyPheProLeuProThrProAlaArgVal                              420425430                                                                     GlnLeuTyrAsnValValLeuGlnProHisGlnAsnPheLeuLeuPhe                              435440445                                                                     GlyAlaAspValValTyrLys                                                         450455                                                                        (2) INFORMATION FOR SEQ ID NO:70:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.74"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:70:                                      LysSerLysValGlyTrpLeuIleGlnLeuPheHisLysLysLysTrp                              151015                                                                        LysAlaGlnLysArgPheLeuLys                                                      20                                                                            (2) INFORMATION FOR SEQ ID NO:71:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.76"                                               (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 10..12                                                          (D) OTHER INFORMATION: /label=D- Phe                                          /note="The amino acid at position 11 is                                       D- phenylalanine"                                                             (xi) SEQUENCE DESCRIPTION: SEQ ID NO:71:                                      IleLysIleSerGlyLysTrpLysAlaGlnPheArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:72:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.77"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:72:                                      IleLysIleSerGlyLysTrpLysAlaGlnTrpArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:73:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.79"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:73:                                      IleLysIleSerGlyLysTrpLysAlaLysLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:74:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.80"                                               (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 10..12                                                          (D) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 11 is                                          beta-1- naphthyl-substituted"                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:74:                                      IleLysIleSerGlyLysTrpLysAlaGlnAlaArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:75:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.81"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:75:                                      IleLysIleSerGlyLysTrpLysAlaPheLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:76:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.82"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:76:                                      LysSerLysValGlyTrpLeuIleGlnLeuTrpHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:77:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.83"                                               (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 10..12                                                          (D) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 6 is                                           beta-1- naphthyl-substituted"                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:77:                                      LysSerLysValGlyAlaLysIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:78:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.84"                                               (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6..8                                                            (D) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 7 is                                           beta-1- naphthyl-substituted"                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:78:                                      IleLysIleSerGlyLysAlaLysAlaGlnPheArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:79:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.85"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:79:                                      LysSerLysValLeuTrpLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:80:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.86"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:80:                                      LysSerLysValGlyTrpLeuIleLeuLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:81:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.87"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:81:                                      LysSerLysValGlyTrpLeuIleGlnLeuPheLeuLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:82:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.88"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:82:                                      IleLysIleSerGlyLysTrpLysAlaPhePheArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:83:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.98"                                               (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 2                                                               (D) OTHER INFORMATION: /label=Substituted-Trp                                 /note="The alanine at position 2 is                                           beta-1- naphthyl-substituted"                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:83:                                      LysTrpLysAlaGlnPheArgPheLeuLysLysSerLysValGlyTrp                              151015                                                                        LeuIlePheLeuPheHisLysLys                                                      20                                                                            (2) INFORMATION FOR SEQ ID NO:84:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.89"                                               (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6..8                                                            (D) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 7 is                                           beta-1- naphthyl-substituted"                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:84:                                      IleLysIleSerGlyLysAlaLysAlaPheLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:85:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.90"                                               (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6..8                                                            (D) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 7 is                                           beta-1- naphthyl-substituted"                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:85:                                      IleLysIleSerGlyLysAlaLysAlaPhePheArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:86:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.91"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:86:                                      LysSerLysValGlyTrpLeuIlePheLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:87:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.92"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:87:                                      LysSerLysValGlyTrpLeuIleLysLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:88:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 29 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.93"                                               (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6..8                                                            (D) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 7 is                                           beta-1- naphthyl-substituted"                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:88:                                      IleLysIleSerGlyLysAlaLysAlaGlnPheArgPheLeuLysLys                              151015                                                                        SerLysValGlyTrpLeuIleGlnLeuPheHisLysLys                                       2025                                                                          (2) INFORMATION FOR SEQ ID NO:89:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.94"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:89:                                      LysSerLysValGlyTrpLeuIleGlnLeuPhePheLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:90:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.95"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:90:                                      LysSerLysValPheTrpLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:91:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.96"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:91:                                      LysSerLysValGlyTrpLeuIleGlnLeuPheHisLysPhe                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:92:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.97"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:92:                                      LysSerLysValLysTrpLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:93:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 30 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.99"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:93:                                      LysTrpLysAlaGlnTrpArgPheLeuLysLysTrpLysAlaGlnTrp                              151015                                                                        ArgPheLeuLysLysTrpLysAlaGlnTrpArgPheLeuLys                                    202530                                                                        (2) INFORMATION FOR SEQ ID NO:94:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.100"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:94:                                      LysSerLysValLysTrpLeuIleLysLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:95:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 28 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.101"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:95:                                      LysSerLysValLysTrpLeuIleLysLeuPhePheLysPheLysSer                              151015                                                                        LysValLysTrpLeuIleLysLeuPhePheLysPhe                                          2025                                                                          (2) INFORMATION FOR SEQ ID NO:96:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.102"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:96:                                      LysTrpLysAlaGlnPheArgPheLeuLysLysSerLysValGlyTrp                              151015                                                                        LeuIleLeuLeuPheHisLysLys                                                      20                                                                            (2) INFORMATION FOR SEQ ID NO:97:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1443 base pairs                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA                                                       (ix) FEATURE:                                                                 (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..1443                                                         (ix) FEATURE:                                                                 (A) NAME/KEY: mat.sub.-- peptide                                              (B) LOCATION: 76..1443                                                        (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "rLBP"                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:97:                                      ATGGGGGCCTTGGCCAGAGCCCTGCCGTCCATACTGCTGGCATTGCTG48                            MetGlyAlaLeuAlaArgAlaLeuProSerIleLeuLeuAlaLeuLeu                              25-20-15-10                                                                   CTTACGTCCACCCCAGAGGCTCTGGGTGCCAACCCCGGCTTGGTCGCC96                            LeuThrSerThrProGluAlaLeuGlyAlaAsnProGlyLeuValAla                              515                                                                           AGGATCACCGACAAGGGACTGCAGTATGCGGCCCAGGAGGGGCTATTG144                           ArgIleThrAspLysGlyLeuGlnTyrAlaAlaGlnGluGlyLeuLeu                              101520                                                                        GCTCTGCAGAGTGAGCTGCTCAGGATCACGCTGCCTGACTTCACCGGG192                           AlaLeuGlnSerGluLeuLeuArgIleThrLeuProAspPheThrGly                              253035                                                                        GACTTGAGGATCCCCCACGTCGGCCGTGGGCGCTATGAGTTCCACAGC240                           AspLeuArgIleProHisValGlyArgGlyArgTyrGluPheHisSer                              40455055                                                                      CTGAACATCCACAGCTGTGAGCTGCTTCACTCTGCGCTGAGGCCTGTC288                           LeuAsnIleHisSerCysGluLeuLeuHisSerAlaLeuArgProVal                              606570                                                                        CCTGGCCAGGGCCTGAGTCTCAGCATCTCCGACTCCTCCATCCGGGTC336                           ProGlyGlnGlyLeuSerLeuSerIleSerAspSerSerIleArgVal                              758085                                                                        CAGGGCAGGTGGAAGGTGCGCAAGTCATTCTTCAAACTACAGGGCTCC384                           GlnGlyArgTrpLysValArgLysSerPhePheLysLeuGlnGlySer                              9095100                                                                       TTTGATGTCAGTGTCAAGGGCATCAGCATTTCGGTCAACCTCCTGTTG432                           PheAspValSerValLysGlyIleSerIleSerValAsnLeuLeuLeu                              105110115                                                                     GGCAGCGAGTCCTCCGGGAGGCCCACAGTTACTGCCTCCAGCTGCAGC480                           GlySerGluSerSerGlyArgProThrValThrAlaSerSerCysSer                              120125130135                                                                  AGTGACATCGCTGACGTGGAGGTGGACATGTCGGGAGACTTGGGGTGG528                           SerAspIleAlaAspValGluValAspMetSerGlyAspLeuGlyTrp                              140145150                                                                     CTGTTGAACCTCTTCCACAACCAGATTGAGTCCAAGTTCCAGAAAGTA576                           LeuLeuAsnLeuPheHisAsnGlnIleGluSerLysPheGlnLysVal                              155160165                                                                     CTGGAGAGCAGGATTTGCGAAATGATCCAGAAATCGGTGTCCTCCGAT624                           LeuGluSerArgIleCysGluMetIleGlnLysSerValSerSerAsp                              170175180                                                                     CTACAGCCTTATCTCCAAACTCTGCCAGTTACAACAGAGATTGACAGT672                           LeuGlnProTyrLeuGlnThrLeuProValThrThrGluIleAspSer                              185190195                                                                     TTCGCCGACATTGATTATAGCTTAGTGGAAGCCCCTCGGGCAACAGCC720                           PheAlaAspIleAspTyrSerLeuValGluAlaProArgAlaThrAla                              200205210215                                                                  CAGATGCTGGAGGTGATGTTTAAGGGTGAAATCTTTCATCGTAACCAC768                           GlnMetLeuGluValMetPheLysGlyGluIlePheHisArgAsnHis                              220225230                                                                     CGTTCTCCAGTTACCCTCCTTGCTGCAGTCATGAGCCTTCCTGAGGAA816                           ArgSerProValThrLeuLeuAlaAlaValMetSerLeuProGluGlu                              235240245                                                                     CACAACAAAATGGTCTACTTTGCCATCTCGGATTATGTCTTCAACACG864                           HisAsnLysMetValTyrPheAlaIleSerAspTyrValPheAsnThr                              250255260                                                                     GCCAGCCTGGTTTATCATGAGGAAGGATATCTGAACTTCTCCATCACA912                           AlaSerLeuValTyrHisGluGluGlyTyrLeuAsnPheSerIleThr                              265270275                                                                     GATGAGATGATACCGCCTGACTCTAATATCCGACTGACCACCAAGTCC960                           AspGluMetIleProProAspSerAsnIleArgLeuThrThrLysSer                              280285290295                                                                  TTCCGACCCTTCGTCCCACGGTTAGCCAGGCTCTACCCCAACATGAAC1008                          PheArgProPheValProArgLeuAlaArgLeuTyrProAsnMetAsn                              300305310                                                                     CTGGAACTCCAGGGATCAGTGCCCTCTGCTCCGCTCCTGAACTTCAGC1056                          LeuGluLeuGlnGlySerValProSerAlaProLeuLeuAsnPheSer                              315320325                                                                     CCTGGGAATCTGTCTGTGGACCCCTATATGGAGATAGATGCCTTTGTG1104                          ProGlyAsnLeuSerValAspProTyrMetGluIleAspAlaPheVal                              330335340                                                                     CTCCTGCCCAGCTCCAGCAAGGAGCCTGTCTTCCGGCTCAGTGTGGCC1152                          LeuLeuProSerSerSerLysGluProValPheArgLeuSerValAla                              345350355                                                                     ACTAATGTGTCCGCCACCTTGACCTTCAATACCAGCAAGATCACTGGG1200                          ThrAsnValSerAlaThrLeuThrPheAsnThrSerLysIleThrGly                              360365370375                                                                  TTCCTGAAGCCAGGAAAGGTAAAAGTGGAACTGAAAGAATCCAAAGTT1248                          PheLeuLysProGlyLysValLysValGluLeuLysGluSerLysVal                              380385390                                                                     GGACTATTCAATGCAGAGCTGTTGGAAGCGCTCCTCAACTATTACATC1296                          GlyLeuPheAsnAlaGluLeuLeuGluAlaLeuLeuAsnTyrTyrIle                              395400405                                                                     CTTAACACCTTCTACCCCAAGTTCAATGATAAGTTGGCCGAAGGCTTC1344                          LeuAsnThrPheTyrProLysPheAsnAspLysLeuAlaGluGlyPhe                              410415420                                                                     CCCCTTCCTCTGCTGAAGCGTGTTCAGCTCTACGACCTTGGGCTGCAG1392                          ProLeuProLeuLeuLysArgValGlnLeuTyrAspLeuGlyLeuGln                              425430435                                                                     ATCCATAAGGACTTCCTGTTCTTGGGTGCCAATGTCCAATACATGAGA1440                          IleHisLysAspPheLeuPheLeuGlyAlaAsnValGlnTyrMetArg                              440445450455                                                                  GTT1443                                                                       Val                                                                           (2) INFORMATION FOR SEQ ID NO:98:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 481 amino acids                                                   (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "rLBP"                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:98:                                      MetGlyAlaLeuAlaArgAlaLeuProSerIleLeuLeuAlaLeuLeu                              25-20-15-10                                                                   LeuThrSerThrProGluAlaLeuGlyAlaAsnProGlyLeuValAla                              515                                                                           ArgIleThrAspLysGlyLeuGlnTyrAlaAlaGlnGluGlyLeuLeu                              101520                                                                        AlaLeuGlnSerGluLeuLeuArgIleThrLeuProAspPheThrGly                              253035                                                                        AspLeuArgIleProHisValGlyArgGlyArgTyrGluPheHisSer                              40455055                                                                      LeuAsnIleHisSerCysGluLeuLeuHisSerAlaLeuArgProVal                              606570                                                                        ProGlyGlnGlyLeuSerLeuSerIleSerAspSerSerIleArgVal                              758085                                                                        GlnGlyArgTrpLysValArgLysSerPhePheLysLeuGlnGlySer                              9095100                                                                       PheAspValSerValLysGlyIleSerIleSerValAsnLeuLeuLeu                              105110115                                                                     GlySerGluSerSerGlyArgProThrValThrAlaSerSerCysSer                              120125130135                                                                  SerAspIleAlaAspValGluValAspMetSerGlyAspLeuGlyTrp                              140145150                                                                     LeuLeuAsnLeuPheHisAsnGlnIleGluSerLysPheGlnLysVal                              155160165                                                                     LeuGluSerArgIleCysGluMetIleGlnLysSerValSerSerAsp                              170175180                                                                     LeuGlnProTyrLeuGlnThrLeuProValThrThrGluIleAspSer                              185190195                                                                     PheAlaAspIleAspTyrSerLeuValGluAlaProArgAlaThrAla                              200205210215                                                                  GlnMetLeuGluValMetPheLysGlyGluIlePheHisArgAsnHis                              220225230                                                                     ArgSerProValThrLeuLeuAlaAlaValMetSerLeuProGluGlu                              235240245                                                                     HisAsnLysMetValTyrPheAlaIleSerAspTyrValPheAsnThr                              250255260                                                                     AlaSerLeuValTyrHisGluGluGlyTyrLeuAsnPheSerIleThr                              265270275                                                                     AspGluMetIleProProAspSerAsnIleArgLeuThrThrLysSer                              280285290295                                                                  PheArgProPheValProArgLeuAlaArgLeuTyrProAsnMetAsn                              300305310                                                                     LeuGluLeuGlnGlySerValProSerAlaProLeuLeuAsnPheSer                              315320325                                                                     ProGlyAsnLeuSerValAspProTyrMetGluIleAspAlaPheVal                              330335340                                                                     LeuLeuProSerSerSerLysGluProValPheArgLeuSerValAla                              345350355                                                                     ThrAsnValSerAlaThrLeuThrPheAsnThrSerLysIleThrGly                              360365370375                                                                  PheLeuLysProGlyLysValLysValGluLeuLysGluSerLysVal                              380385390                                                                     GlyLeuPheAsnAlaGluLeuLeuGluAlaLeuLeuAsnTyrTyrIle                              395400405                                                                     LeuAsnThrPheTyrProLysPheAsnAspLysLeuAlaGluGlyPhe                              410415420                                                                     ProLeuProLeuLeuLysArgValGlnLeuTyrAspLeuGlyLeuGln                              425430435                                                                     IleHisLysAspPheLeuPheLeuGlyAlaAsnValGlnTyrMetArg                              440445450455                                                                  Val                                                                           (2) INFORMATION FOR SEQ ID NO:99:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 16 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.57"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:99:                                      CysIleLysIleSerGlyLysTrpLysAlaGlnLysArgProLeuCys                              151015                                                                        (2) INFORMATION FOR SEQ ID NO:100:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.75"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:100:                                     IleLysLysArgAlaIleSerPheLeuGlyLysLysTrpGlnLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:101:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.282"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:101:                                     LysTrpLysAlaPhePheArgPheLeuLysLysTrpLysAlaPhePhe                              151015                                                                        ArgPheLeuLys                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:102:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 16 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.103"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:102:                                     IleLysIleSerGlyLysTrpLysAlaTrpLysArgPheLeuLysLys                              151015                                                                        (2) INFORMATION FOR SEQ ID NO:103:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.104"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:103:                                     LysSerLysValGlyTrpLeuIleSerLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:104:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 16 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.105"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 13                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 13 is beta-1-                                  naphthyl- substituted."                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO:104:                                     IleLysIleSerGlyLysTrpLysAlaTrpLysArgAlaLeuLysLys                              151015                                                                        (2) INFORMATION FOR SEQ ID NO:105:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.106"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:105:                                     LysSerLysValGlyTrpLeuIleThrLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:106:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.107"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:106:                                     LysSerLysValGlyTrpLeuIleGlnLeuPheTrpLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:107:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.108"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:107:                                     LysSerLysValGlyTrpLeuIleGlnLeuPheHisLysTrp                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:108:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.109"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 11                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 11 is beta-1-                                  naphthyl- substituted."                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO:108:                                     LysSerLysValGlyTrpLeuIleGlnLeuAlaHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:109:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.110"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 12                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 12 is beta-1-                                  naphthyl- substituted."                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO:109:                                     LysSerLysValGlyTrpLeuIleGlnLeuPheAlaLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:110:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.111"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 14                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 14 is beta-1-                                  naphthyl- substituted."                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO:110:                                     LysSerLysValGlyTrpLeuIleGlnLeuPheHisLysAla                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:111:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.112"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 7                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 7 is beta-1-                                   naphthyl- substituted."                                                       (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 11                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 11 is beta-1-                                  naphthyl- substituted."                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO:111:                                     IleLysIleSerGlyLysAlaLysAlaGlnAlaArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:112:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.113"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:112:                                     LysSerLysValGlyTrpLeuIleGlnPhePheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:113:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.114"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:113:                                     LysTrpGlnLeuArgSerLysGlyLysIleLysIlePheLysAla                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:114:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.116"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 6 is beta-1-                                   naphthyl- substituted."                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO:114:                                     LysSerLysValLysAlaLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:115:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.119"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 7                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 7 is beta-1-                                   naphthyl- substituted."                                                       (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 10                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 10 is beta-1-                                  naphthyl- substituted."                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO:115:                                     IleLysIleSerGlyLysAlaLysAlaAlaLysArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:116:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.120"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:116:                                     IleLysIleSerGlyLysTrpLysAlaGlnLysArgLysLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:117:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.121"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 10                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 10 is beta-1-                                  naphthyl- substituted."                                                       (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 11                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 11 is beta-1-                                  naphthyl- substituted."                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO:117:                                     IleLysIleSerGlyLysTrpLysAlaAlaAlaArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:118:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.122"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 7                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 7 is beta-1-                                   naphthyl- substituted."                                                       (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 10                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 10 is beta-1-                                  naphthyl- substituted."                                                       (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 11                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 11 is beta-1-                                  naphthyl- substituted."                                                       (xi) SEQUENCE DESCRIPTION: SEQ ID NO:118:                                     IleLysIleSerGlyLysAlaLysAlaAlaAlaArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:119:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.123"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 9                                                               (C) OTHER INFORMATION: /label=Substituted-Phe                                 /note="The phenylalanine at position 9 is                                     p-amino- substituted."                                                        (xi) SEQUENCE DESCRIPTION: SEQ ID NO:119:                                     LysSerLysValGlyTrpLeuIlePheLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:120:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.124"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:120:                                     LysSerLysValLysTrpLeuIleGlnLeuTrpHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:121:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.125"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:121:                                     LysSerLysValGlyTrpLeuIleTyrLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:122:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.126"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=D- Trp                                          /note="The amino acid at position 6 is                                        D- tryptophan."                                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:122:                                     LysSerLysValGlyTrpLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:123:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.127"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:123:                                     LysSerLysValGlyPheLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:124:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.128"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=D- Phe                                          /note="The amino acid at position 6 is                                        D- phenylalanine."                                                            (xi) SEQUENCE DESCRIPTION: SEQ ID NO:124:                                     LysSerLysValGlyPheLeuIleGlnLeuProHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:125:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.129"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 6 is                                           D-1-beta-1- naphthyl-                                                         substituted."                                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:125:                                     LysSerLysValGlyAlaLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:126:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.130"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 6 is                                           2-beta-1- naphthyl-                                                           substituted."                                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:126:                                     LysSerLysValGlyAlaLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:127:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.131"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 6 is                                           D-2-beta-1- naphthyl-                                                         substituted."                                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:127:                                     LysSerLysValGlyAlaLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:128:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.132"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 6 is                                           pyridyl- substituted."                                                        (xi) SEQUENCE DESCRIPTION: SEQ ID NO:128:                                     LysSerLysValGlyAlaLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:129:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.133"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Phe                                 /note="The phenylalanine at position 6 is                                     para-amino-                                                                   substituted."                                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:129:                                     LysSerLysValGlyPheLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:130:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.134"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 5                                                               (C) OTHER INFORMATION: /label=Substituted-Phe                                 /note="The phenylalanine at position 5 is                                     para-amino- substituted."                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:130:                                     LysSerLysValPheTrpLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:131:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.135"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:131:                                     LysSerLysValGlyLysLeuIleGlnLeuProHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:132:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.136"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:132:                                     IleLysIleSerGlyLysTrpLysAlaGlnGluArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:133:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 16 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.137"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:133:                                     CysLysSerLysValGlyTrpLeuIleGlnLeuPheHisLysLysCys                              151015                                                                        (2) INFORMATION FOR SEQ ID NO:134:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.138"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:134:                                     LysSerLysValLysPheLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:135:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.139"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:135:                                     LysSerLysValGlyTyrLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:136:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.140"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 1                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 1 is                                           beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 2                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 2 is                                           beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:136:                                     AlaAlaArgPheLeuLysPhe                                                         15                                                                            (2) INFORMATION FOR SEQ ID NO:137:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.141"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:137:                                     IleLysIleSerGlyLysTrpLysAlaGlnLysArgTrpLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:138:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.142"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:138:                                     LysSerLysValGlyTrpLeuIleGlnTrpPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:139:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.143"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 10                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 10 is                                          beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:139:                                     LysSerLysValGlyTrpLeuIleGlnAlaPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:140:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.144"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 6 is                                           cyclohexyl- substituted."                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:140:                                     LysSerLysValGlyAlaLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:141:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.145"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:141:                                     LysTrpLysAlaAlaAlaArgPheLeuLysLysSerLysValGlyTrp                              151015                                                                        LeuIleGlnLeuPheHisLysLys                                                      20                                                                            (2) INFORMATION FOR SEQ ID NO:142:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.146"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 12                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 12 is                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 14                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 14 is                                          beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:142:                                     LysSerLysValGlyTrpLeuIleGlnLeuPheAlaLysAla                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:143:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.147"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:143:                                     IleLysIleSerGlyLysTrpLysAlaGluLysLysPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:144:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.148"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 6 is                                           beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 12                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 12 is                                          beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:144:                                     LysSerLysValGlyAlaLeuIleGlnLeuPheAlaLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:145:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1813 base pairs                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (ix) FEATURE:                                                                 (A) NAME/KEY: CDS                                                             (B) LOCATION: 31..1491                                                        (ix) FEATURE:                                                                 (A) NAME/KEY: mat.sub.-- peptide                                              (B) LOCATION: 124..1491                                                       (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "rBPI"                                                 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:145:                                     CAGGCCTTGAGGTTTTGGCAGCTCTGGAGGATGAGAGAGAACATGGCCAGGGGC54                      MetArgGluAsnMetAlaArgGly                                                      31-30-25                                                                      CCTTGCAACGCGCCGAGATGGGTGTCCCTGATGGTGCTCGTCGCCATA102                           ProCysAsnAlaProArgTrpValSerLeuMetValLeuValAlaIle                              20-15- 10                                                                     GGCACCGCCGTGACAGCGGCCGTCAACCCTGGCGTCGTGGTCAGGATC150                           GlyThrAlaValThrAlaAlaValAsnProGlyValValValArgIle                              515                                                                           TCCCAGAAGGGCCTGGACTACGCCAGCCAGCAGGGGACGGCCGCTCTG198                           SerGlnLysGlyLeuAspTyrAlaSerGlnGlnGlyThrAlaAlaLeu                              10152025                                                                      CAGAAGGAGCTGAAGAGGATCAAGATTCCTGACTACTCAGACAGCTTT246                           GlnLysGluLeuLysArgIleLysIleProAspTyrSerAspSerPhe                              303540                                                                        AAGATCAAGCATCTTGGGAAGGGGCATTATAGCTTCTACAGCATGGAC294                           LysIleLysHisLeuGlyLysGlyHisTyrSerPheTyrSerMetAsp                              455055                                                                        ATCCGTGAATTCCAGCTTCCCAGTTCCCAGATAAGCATGGTGCCCAAT342                           IleArgGluPheGlnLeuProSerSerGlnIleSerMetValProAsn                              606570                                                                        GTGGGCCTTAAGTTCTCCATCAGCAACGCCAATATCAAGATCAGCGGG390                           ValGlyLeuLysPheSerIleSerAsnAlaAsnIleLysIleSerGly                              758085                                                                        AAATGGAAGGCACAAAAGAGATTCTTAAAAATGAGCGGCAATTTTGAC438                           LysTrpLysAlaGlnLysArgPheLeuLysMetSerGlyAsnPheAsp                              9095100105                                                                    CTGAGCATAGAAGGCATGTCCATTTCGGCTGATCTGAAGCTGGGCAGT486                           LeuSerIleGluGlyMetSerIleSerAlaAspLeuLysLeuGlySer                              110115120                                                                     AACCCCACGTCAGGCAAGCCCACCATCACCTGCTCCAGCTGCAGCAGC534                           AsnProThrSerGlyLysProThrIleThrCysSerSerCysSerSer                              125130135                                                                     CACATCAACAGTGTCCACGTGCACATCTCAAAGAGCAAAGTCGGGTGG582                           HisIleAsnSerValHisValHisIleSerLysSerLysValGlyTrp                              140145150                                                                     CTGATCCAACTCTTCCACAAAAAAATTGAGTCTGCGCTTCGAAACAAG630                           LeuIleGlnLeuPheHisLysLysIleGluSerAlaLeuArgAsnLys                              155160165                                                                     ATGAACAGCCAGGTCTGCGAGAAAGTGACCAATTCTGTATCCTCCAAG678                           MetAsnSerGlnValCysGluLysValThrAsnSerValSerSerLys                              170175180185                                                                  CTGCAACCTTATTTCCAGACTCTGCCAGTAATGACCAAAATAGATTCT726                           LeuGlnProTyrPheGlnThrLeuProValMetThrLysIleAspSer                              190195200                                                                     GTGGCTGGAATCAACTATGGTCTGGTGGCACCTCCAGCAACCACGGCT774                           ValAlaGlyIleAsnTyrGlyLeuValAlaProProAlaThrThrAla                              205210215                                                                     GAGACCCTGGATGTACAGATGAAGGGGGAGTTTTACAGTGAGAACCAC822                           GluThrLeuAspValGlnMetLysGlyGluPheTyrSerGluAsnHis                              220225230                                                                     CACAATCCACCTCCCTTTGCTCCACCAGTGATGGAGTTTCCCGCTGCC870                           HisAsnProProProPheAlaProProValMetGluPheProAlaAla                              235240245                                                                     CATGACCGCATGGTATACCTGGGCCTCTCAGACTACTTCTTCAACACA918                           HisAspArgMetValTyrLeuGlyLeuSerAspTyrPhePheAsnThr                              250255260265                                                                  GCCGGGCTTGTATACCAAGAGGCTGGGGTCTTGAAGATGACCCTTAGA966                           AlaGlyLeuValTyrGlnGluAlaGlyValLeuLysMetThrLeuArg                              270275280                                                                     GATGACATGATTCCAAAGGAGTCCAAATTTCGACTGACAACCAAGTTC1014                          AspAspMetIleProLysGluSerLysPheArgLeuThrThrLysPhe                              285290295                                                                     TTTGGAACCTTCCTACCTGAGGTGGCCAAGAAGTTTCCCAACATGAAG1062                          PheGlyThrPheLeuProGluValAlaLysLysPheProAsnMetLys                              300305310                                                                     ATACAGATCCATGTCTCAGCCTCCACCCCGCCACACCTGTCTGTGCAG1110                          IleGlnIleHisValSerAlaSerThrProProHisLeuSerValGln                              315320325                                                                     CCCACCGGCCTTACCTTCTACCCTGCCGTGGATGTCCAGGCCTTTGCC1158                          ProThrGlyLeuThrPheTyrProAlaValAspValGlnAlaPheAla                              330335340345                                                                  GTCCTCCCCAACTCCTCCCTGGCTTCCCTCTTCCTGATTGGCATGCAC1206                          ValLeuProAsnSerSerLeuAlaSerLeuPheLeuIleGlyMetHis                              350355360                                                                     ACAACTGGTTCCATGGAGGTCAGCGCCGAGTCCAACAGGCTTGTTGGA1254                          ThrThrGlySerMetGluValSerAlaGluSerAsnArgLeuValGly                              365370375                                                                     GAGCTCAAGCTGGATAGGCTGCTCCTGGAACTGAAGCACTCAAATATT1302                          GluLeuLysLeuAspArgLeuLeuLeuGluLeuLysHisSerAsnIle                              380385390                                                                     GGCCCCTTCCCGGTTGAATTGCTGCAGGATATCATGAACTACATTGTA1350                          GlyProPheProValGluLeuLeuGlnAspIleMetAsnTyrIleVal                              395400405                                                                     CCCATTCTTGTGCTGCCCAGGGTTAACGAGAAACTACAGAAAGGCTTC1398                          ProIleLeuValLeuProArgValAsnGluLysLeuGlnLysGlyPhe                              410415420425                                                                  CCTCTCCCGACGCCGGCCAGAGTCCAGCTCTACAACGTAGTGCTTCAG1446                          ProLeuProThrProAlaArgValGlnLeuTyrAsnValValLeuGln                              430435440                                                                     CCTCACCAGAACTTCCTGCTGTTCGGTGCAGACGTTGTCTATAAA1491                             ProHisGlnAsnPheLeuLeuPheGlyAlaAspValValTyrLys                                 445450455                                                                     TGAAGGCACCAGGGGTGCCGGGGGCTGTCAGCCGCACCTGTTCCTGATGGGCTGTGGGGC1551              ACCGGCTGCCTTTCCCCAGGGAATCCTCTCCAGATCTTAACCAAGAGCCCCTTGCAAACT1611              TCTTCGACTCAGATTCAGAAATGATCTAAACACGAGGAAACATTATTCATTGGAAAAGTG1671              CATGGTGTGTATTTTAGGGATTATGAGCTTCTTTCAAGGGCTAAGGCTGCAGAGATATTT1731              CCTCCAGGAATCGTGTTTCAATTGTAACCAAGAAATTTCCATTTGTGCTTCATGAAAAAA1791              AACTTCTGGTTTTTTTCATGTG1813                                                    (2) INFORMATION FOR SEQ ID NO:146:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 487 amino acids                                                   (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:146:                                     MetArgGluAsnMetAlaArgGlyProCysAsnAlaProArgTrpVal                              31-30-25-20                                                                   SerLeuMetValLeuValAlaIleGlyThrAlaValThrAlaAlaVal                              15-10-51                                                                      AsnProGlyValValValArgIleSerGlnLysGlyLeuAspTyrAla                              51015                                                                         SerGlnGlnGlyThrAlaAlaLeuGlnLysGluLeuLysArgIleLys                              202530                                                                        IleProAspTyrSerAspSerPheLysIleLysHisLeuGlyLysGly                              354045                                                                        HisTyrSerPheTyrSerMetAspIleArgGluPheGlnLeuProSer                              50556065                                                                      SerGlnIleSerMetValProAsnValGlyLeuLysPheSerIleSer                              707580                                                                        AsnAlaAsnIleLysIleSerGlyLysTrpLysAlaGlnLysArgPhe                              859095                                                                        LeuLysMetSerGlyAsnPheAspLeuSerIleGluGlyMetSerIle                              100105110                                                                     SerAlaAspLeuLysLeuGlySerAsnProThrSerGlyLysProThr                              115120125                                                                     IleThrCysSerSerCysSerSerHisIleAsnSerValHisValHis                              130135140145                                                                  IleSerLysSerLysValGlyTrpLeuIleGlnLeuPheHisLysLys                              150155160                                                                     IleGluSerAlaLeuArgAsnLysMetAsnSerGlnValCysGluLys                              165170175                                                                     ValThrAsnSerValSerSerLysLeuGlnProTyrPheGlnThrLeu                              180185190                                                                     ProValMetThrLysIleAspSerValAlaGlyIleAsnTyrGlyLeu                              195200205                                                                     ValAlaProProAlaThrThrAlaGluThrLeuAspValGlnMetLys                              210215220225                                                                  GlyGluPheTyrSerGluAsnHisHisAsnProProProPheAlaPro                              230235240                                                                     ProValMetGluPheProAlaAlaHisAspArgMetValTyrLeuGly                              245250255                                                                     LeuSerAspTyrPhePheAsnThrAlaGlyLeuValTyrGlnGluAla                              260265270                                                                     GlyValLeuLysMetThrLeuArgAspAspMetIleProLysGluSer                              275280285                                                                     LysPheArgLeuThrThrLysPhePheGlyThrPheLeuProGluVal                              290295300305                                                                  AlaLysLysPheProAsnMetLysIleGlnIleHisValSerAlaSer                              310315320                                                                     ThrProProHisLeuSerValGlnProThrGlyLeuThrPheTyrPro                              325330335                                                                     AlaValAspValGlnAlaPheAlaValLeuProAsnSerSerLeuAla                              340345350                                                                     SerLeuPheLeuIleGlyMetHisThrThrGlySerMetGluValSer                              355360365                                                                     AlaGluSerAsnArgLeuValGlyGluLeuLysLeuAspArgLeuLeu                              370375380385                                                                  LeuGluLeuLysHisSerAsnIleGlyProPheProValGluLeuLeu                              390395400                                                                     GlnAspIleMetAsnTyrIleValProIleLeuValLeuProArgVal                              405410415                                                                     AsnGluLysLeuGlnLysGlyPheProLeuProThrProAlaArgVal                              420425430                                                                     GlnLeuTyrAsnValValLeuGlnProHisGlnAsnPheLeuLeuPhe                              435440445                                                                     GlyAlaAspValValTyrLys                                                         450455                                                                        (2) INFORMATION FOR SEQ ID NO:147:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.149"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:147:                                     LysTrpLysValPheLysLysIleGluLysLysSerLysValGlyTrp                              151015                                                                        LeuIleGlnLeuPheHisLysLys                                                      20                                                                            (2) INFORMATION FOR SEQ ID NO:148:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.150"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:148:                                     LysTrpAlaPheAlaLysLysGlnLysLysArgLeuLysArgGlnTrp                              151015                                                                        LeuLysLysPhe                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:149:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 30 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.153"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:149:                                     LysTrpLysAlaGlnLysArgPheLeuLysLysTrpLysAlaGlnLys                              151015                                                                        ArgPheLeuLysLysTrpLysAlaGlnLysArgPheLeuLys                                    202530                                                                        (2) INFORMATION FOR SEQ ID NO:150:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.154"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 5                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 5 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 6 is                                                          beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:150:                                     LysTrpLysAlaAlaAlaArgPheLeuLysLysTrpLysAlaGlnLys                              151015                                                                        ArgPheLeuLys                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:151:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.155"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 15                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 15 is                                                         beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 16                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 16 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:151:                                     LysTrpLysAlaGlnLysArgPheLeuLysLysTrpLysAlaAlaAla                              151015                                                                        ArgPheLeuLys                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:152:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.156"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 5                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 5 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 6 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 15                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 15 is                                                         beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 16                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 16 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:152:                                     LysTrpLysAlaAlaAlaArgPheLeuLysLysTrpLysAlaAlaAla                              151015                                                                        ArgPheLeuLys                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:153:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 30 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.157"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 5                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 5 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 6 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 15                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 15 is                                                         beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 16                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 16 is                                                         beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 25                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 25 is                                                         beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 26                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 26 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:153:                                     LysTrpLysAlaAlaAlaArgPheLeuLysLysTrpLysAlaAlaAla                              151015                                                                        ArgPheLeuLysLysTrpLysAlaAlaAlaArgPheLeuLys                                    202530                                                                        (2) INFORMATION FOR SEQ ID NO:154:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 29 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.158"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 10                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 10 is                                                         beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 11                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 11 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:154:                                     IleLysIleSerGlyLysTrpLysAlaAlaAlaArgPheLeuLysLys                              151015                                                                        SerLysValGlyTrpLeuIleGlnLeuPheHisLysLys                                       2025                                                                          (2) INFORMATION FOR SEQ ID NO:155:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.159"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 2                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 2 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 6 is                                                          beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:155:                                     LysAlaLysAlaGlnAlaArgPheLeuLysLysSerLysValGlyTrp                              151015                                                                        LeuIleGlnLeuTrpHisLysLys                                                      20                                                                            (2) INFORMATION FOR SEQ ID NO:156:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.160"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 2                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 2 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 6 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 12                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 12 is                                                         beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 16                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 16 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:156:                                     LysAlaLysAlaGlnAlaArgPheLeuLysLysAlaLysAlaGlnAla                              151015                                                                        ArgPheLeuLys                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:157:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.161"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:157:                                     LysSerLysValLysAlaLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:158:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.162"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:158:                                     LysTrpLysAlaGlnTrpArgPheLeuLysLysSerLysValGlyTrp                              151015                                                                        LeuIleGlnLeuPheHisLysLys                                                      20                                                                            (2) INFORMATION FOR SEQ ID NO:159:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.163"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:159:                                     LysTrpLysAlaGlnTrpArgPheLeuLysLysTrpLysAlaGlnTrp                              151015                                                                        ArgPheLeuLys                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:160:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.164"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 5                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 5 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 15                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 15 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:160:                                     LysTrpLysAlaAlaLysArgPheLeuLysLysTrpLysAlaAlaLys                              151015                                                                        ArgPheLeuLys                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:161:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.165"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 2                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 2 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 12                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 12 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:161:                                     LysAlaLysAlaGlnPheArgPheLeuLysLysAlaLysAlaGlnPhe                              151015                                                                        ArgPheLeuLys                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:162:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.166"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:162:                                     LysSerLysValGlyValLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:163:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 8 amino acids                                                     (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.167"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:163:                                     LysTrpLysAlaGlnLysArgPhe                                                      15                                                                            (2) INFORMATION FOR SEQ ID NO:164:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: circular                                                        (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.168"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:164:                                     CysLysTrpLysAlaGlnLysArgPheLeuLysMetSerCys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:165:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: circular                                                        (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.169"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:165:                                     CysLysTrpLysAlaGlnLysArgPheCys                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:166:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.221"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 13                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 13 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:166:                                     IleLysIleSerGlyLysTrpLysAlaGlnLysArgAlaLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:167:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.222"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 6 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 14                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 14 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:167:                                     LysSerLysValGlyAlaLeuIleGlnLeuPheHisLysAla                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:168:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.223"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 6 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 10                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 10 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:168:                                     LysSerLysValGlyAlaLeuIleGlnAlaPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:169:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.224"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 6 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 9                                                               (C) OTHER INFORMATION: /label=Substituted-Phe                                 /note="Position 9 is                                                          para-amino- substituted."                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:169:                                     LysSerLysValGlyAlaLeuIlePheLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:170:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.225"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 6 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 5                                                               (C) OTHER INFORMATION: /label=Substituted-Phe                                 /note="Position 5 is                                                          para-amino- substituted."                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:170:                                     LysSerLysValPheAlaLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:171:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.226"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 6 is                                                          beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:171:                                     LysSerLysValGlyAlaLeuIleGlnLeuTrpHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:172:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.227"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 10                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 10 is                                                         beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 14                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 14 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:172:                                     LysSerLysValGlyTrpLeuIleGlnAlaPheHisLysAla                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:173:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.228"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 9                                                               (C) OTHER INFORMATION: /label=Substituted-Phe                                 /note="Position 9 is                                                          para-amino- substituted."                                                     (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 14                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 14 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:173:                                     LysSerLysValGlyTrpLeuIlePheLeuPheHisLysAla                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:174:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.229"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 5                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 5 is                                                          para-amino- substituted."                                                     (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 14                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 14 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:174:                                     LysSerLysValPheTrpLeuIleGlnLeuPheHisLysAla                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:175:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.230"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 14                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 14 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:175:                                     LysSerLysValGlyTrpLeuIleGlnLeuTrpHisLysAla                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:176:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.231"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 10                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 10 is                                                         beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 12                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 12 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:176:                                     LysSerLysValGlyTrpLeuIleGlnAlaPheAlaLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:177:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.232"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 9                                                               (C) OTHER INFORMATION: /label=Substituted-Phe                                 /note="Position 9 is                                                          para-amino- substituted."                                                     (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 12                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 12 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:177:                                     LysSerLysValGlyTrpLeuIlePheLeuPheAlaLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:178:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.233"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 5                                                               (C) OTHER INFORMATION: /label=Substituted-Phe                                 /note="Position 5 is                                                          para-amino- substituted."                                                     (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 12                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 12 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:178:                                     LysSerLysValPheTrpLeuIleGlnLeuPheAlaLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:179:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.234"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 12                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 12 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:179:                                     LysSerLysValGlyTrpLeuIleGlnLeuTrpAlaLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:180:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.235"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 9                                                               (C) OTHER INFORMATION: /label=Substituted-Phe                                 /note="Position 9 is                                                          para-amino- substituted."                                                     (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 10                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 10 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:180:                                     LysSerLysValGlyTrpLeuIlePheAlaPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:181:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.236"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 5                                                               (C) OTHER INFORMATION: /label=Substituted-Phe                                 /note="Position 5 is                                                          para-amino- substituted."                                                     (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 10                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 10 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:181:                                     LysSerLysValPheTrpLeuIleGlnAlaPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:182:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.237"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 10                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 10 is                                                         beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:182:                                     LysSerLysValGlyTrpLeuIleGlnAlaTrpHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:183:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.238"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 5                                                               (C) OTHER INFORMATION: /label=Substituted-Phe                                 /note="Position 5 is                                                          para-amino- substituted."                                                     (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 9                                                               (C) OTHER INFORMATION: /label=Substituted-Phe                                 /note="Position 9 is                                                          para-amino- substituted."                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:183:                                     LysSerLysValPheTrpLeuIlePheLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:184:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.239"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 9                                                               (C) OTHER INFORMATION: /label=Substituted-Phe                                 /note="Position 9 is                                                          para-amino- substituted."                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:184:                                     LysSerLysValGlyTrpLeuIlePheLeuTrpHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:185:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.240"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 5                                                               (C) OTHER INFORMATION: /label=Substituted-Phe                                 /note="Position 5 is                                                          para-amino- substituted."                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:185:                                     LysSerLysValPheTrpLeuIleGlnLeuTrpHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:186:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.247"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 2                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 2 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 6 is                                                          beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:186:                                     LysAlaLysAlaGlnAlaArgPheLeuLysLysSerLysValGlyTrp                              151015                                                                        LeuIleLeuLeuPheHisLysLys                                                      20                                                                            (2) INFORMATION FOR SEQ ID NO:187:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.245"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:187:                                     LysTrpLysAlaGlnPheArgPheLeuLysLysSerLysValGlyTrp                              151015                                                                        LeuIleGlnLeuTrpHisLysLys                                                      20                                                                            (2) INFORMATION FOR SEQ ID NO:188:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.246"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 16                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 16 is                                                         D-beta-2- naphthyl-substituted."                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:188:                                     LysTrpLysAlaGlnPheArgPheLeuLysLysSerLysValGlyAla                              151015                                                                        LeuIleGlnLeuPheHisLysLys                                                      20                                                                            (2) INFORMATION FOR SEQ ID NO:189:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.248"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 2                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 2 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 6 is                                                          beta-1- naphthyl-substituted."                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 16                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 16 is                                                         D-beta-2- naphthyl-substituted."                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:189:                                     LysAlaLysAlaGlnAlaArgPheLeuLysLysSerLysValGlyAla                              151015                                                                        LeuIleGlnLeuPheHisLysLys                                                      20                                                                            (2) INFORMATION FOR SEQ ID NO:190:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.242"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 6 is                                                          D-beta-2- naphthyl-substituted."                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:190:                                     LysSerLysValGlyAlaLeuIleLeuLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:191:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 28 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.272"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:191:                                     LysSerLysValGlyTrpLeuIleLeuLeuPheHisLysLysLysSer                              151015                                                                        LysValGlyTrpLeuIleLeuLeuPheHisLysLys                                          2025                                                                          (2) INFORMATION FOR SEQ ID NO:192:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 28 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.275"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:192:                                     LysSerLysValGlyTrpLeuIlePheLeuPheHisLysLysLysSer                              151015                                                                        LysValGlyTrpLeuIlePheLeuPheHisLysLys                                          2025                                                                          (2) INFORMATION FOR SEQ ID NO:193:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 28 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.270"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:193:                                     LysSerLysValGlyTrpLeuIleLeuLeuPheHisLysLysLysSer                              151015                                                                        LysValGlyTrpLeuIleGlnLeuPheHisLysLys                                          2025                                                                          (2) INFORMATION FOR SEQ ID NO:194:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 28 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.271"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:194:                                     LysSerLysValGlyTrpLeuIleGlnLeuPheHisLysLysLysSer                              151015                                                                        LysValGlyTrpLeuIleLeuLeuPheHisLysLys                                          2025                                                                          (2) INFORMATION FOR SEQ ID NO:195:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 28 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.273"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:195:                                     LysSerLysValGlyTrpLeuIlePheLeuPheHisLysLysLysSer                              151015                                                                        LysValGlyTrpLeuIleGlnLeuPheHisLysLys                                          2025                                                                          (2) INFORMATION FOR SEQ ID NO:196:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 28 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.274"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:196:                                     LysSerLysValGlyTrpLeuIleGlnLeuPheHisLysLysLysSer                              151015                                                                        LysValGlyTrpLeuIlePheLeuPheHisLysLys                                          2025                                                                          (2) INFORMATION FOR SEQ ID NO:197:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.276"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:197:                                     LysTrpLysAlaGlnPheArgPheLeuLysLysSerLysValGlyTrp                              151015                                                                        LeuIlePheLeuPheHisLysLys                                                      20                                                                            (2) INFORMATION FOR SEQ ID NO:198:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.241"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:198:                                     LysSerLysValGlyTrpLeuIleLeuLeuTrpHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:199:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.243"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 6 is                                                          D-beta-2- naphthyl-substituted."                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:199:                                     LysSerLysValGlyAlaLeuIleGlnLeuTrpHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:200:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.244"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="Position 6 is                                                          D-beta-2- naphthyl-substituted."                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:200:                                     LysSerLysValGlyAlaLeuIleLeuLeuTrpHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:201:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.249"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:201:                                     LysSerLysValGlyGlyLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:202:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.250"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:202:                                     LysSerLysValGlyLeuLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:203:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.251"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:203:                                     LysSerLysValGlyIleLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:204:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.252"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (D) OTHER INFORMATION: /label=D- Ala                                          /note="The amino acid at position 6 is                                        D-alanine"                                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:204:                                     LysSerLysValGlyAlaLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:205:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.253"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (D) OTHER INFORMATION: /label=D- Val                                          /note="The amino acid at position 6 is                                        D-valine"                                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:205:                                     LysSerLysValGlyValLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:206:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.254"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (D) OTHER INFORMATION: /label=beta-Ala                                        /note="The amino acid at position 6 is                                        beta- alanine"                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:206:                                     LysSerLysValGlyAlaLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:207:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.255"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (D) OTHER INFORMATION: /label=delta-aba                                       /note="The amino acid at position 6 is                                        delta- aminobutyric acid"                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:207:                                     LysSerLysValGlyXaaLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:208:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.256"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (D) OTHER INFORMATION: /label=gaba                                            /note="The amino acid at position 6 is                                        gamma- aminobutyric acid"                                                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:208:                                     LysSerLysValGlyXaaLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:209:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.257"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (D) OTHER INFORMATION: /label=d- methyl-A                                     /note="The amino acid at position 6 is                                        delta-Methyl- alanine"                                                        (xi) SEQUENCE DESCRIPTION: SEQ ID NO:209:                                     LysSerLysValGlyAlaLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:210:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.258"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (D) OTHER INFORMATION: /label=t- butyl-G                                      /note="The amino acid at position 6 is                                        tert-butyl- glycine"                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:210:                                     LysSerLysValGlyGlyLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:211:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.259"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (D) OTHER INFORMATION: /label=N- methyl-G                                     /note="The amino acid at position 6 is                                        N-Methyl- glycine"                                                            (xi) SEQUENCE DESCRIPTION: SEQ ID NO:211:                                     LysSerLysValGlyGlyLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:212:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.260"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (D) OTHER INFORMATION: /label=N- methyl-V                                     /note="The amino acid at position 6 is                                        N-Methyl- valine"                                                             (xi) SEQUENCE DESCRIPTION: SEQ ID NO:212:                                     LysSerLysValGlyValLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:213:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.261"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 6                                                               (D) OTHER INFORMATION: /label=N- methyl-L                                     /note="The amino acid at position 6 is                                        N-Methyl- leucine"                                                            (xi) SEQUENCE DESCRIPTION: SEQ ID NO:213:                                     LysSerLysValGlyLeuLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:214:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.262"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:214:                                     LysSerLysValGlyTrpLeuIleAsnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:215:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.263"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:215:                                     LysSerLysValGlyTrpLeuIleGluLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:216:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.264"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:216:                                     LysSerLysValGlyTrpLeuIleAspLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:217:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.265"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:217:                                     LysSerLysValGlyTrpLeuIleLysLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:218:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.266"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:218:                                     LysSerLysValLysValLeuIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:219:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.267"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:219:                                     LysSerLysValLysTrpAlaIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:220:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.268"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:220:                                     LysSerLysValGlyValAlaIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:221:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.269"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:221:                                     LysSerLysValLysValAlaIleGlnLeuPheHisLysLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:222:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.277"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 2                                                               (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 2 is                                           beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:222:                                     LysAlaLysAlaGlnPheArgPheLeuLysLysSerLysValGlyTrp                              151015                                                                        LeuIleLeuLeuPheHisLysLys                                                      20                                                                            (2) INFORMATION FOR SEQ ID NO:223:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.278"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:223:                                     IleLysIleSerGlyLysTrpLysAlaAlaTrpArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:224:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.279"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 10                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 10 is                                          beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:224:                                     IleLysIleSerGlyLysTrpLysAlaAlaPheArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:225:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.280"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:225:                                     IleLysIleSerGlyLysTrpLysAlaAlaPheArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:226:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.281"                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Modified-site                                                   (B) LOCATION: 10                                                              (C) OTHER INFORMATION: /label=Substituted-Ala                                 /note="The alanine at position 10 is                                          beta-1- naphthyl-substituted."                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:226:                                     IleLysIleSerGlyLysTrpLysAlaAlaAlaArgPheLeuLys                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:227:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "XMP.170"                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:227:                                     LysTrpLysAlaGlnLysArgPheLeuLysMetSer                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:228:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 21 base pairs                                                     (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA                                                       (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "LBP-10"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:228:                                     GAGATCGATAGTTTCGCCGAC21                                                       (2) INFORMATION FOR SEQ ID NO:229:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 base pairs                                                     (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA                                                       (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "LBP-11"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:229:                                     ACTATCGATCTCTGTTGTAA20                                                        (2) INFORMATION FOR SEQ ID NO:230:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 29 base pairs                                                     (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA                                                       (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "LBP- BSM"                                             (xi) SEQUENCE DESCRIPTION: SEQ ID NO:230:                                     GAATGCAGCCAACCCCGGCTTGGTCGCCA29                                               (2) INFORMATION FOR SEQ ID NO:231:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 18 base pairs                                                     (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA                                                       (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "LBP-8"                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:231:                                     CTGGCTAACCGTGGGACG18                                                          (2) INFORMATION FOR SEQ ID NO:232:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 base pairs                                                     (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA                                                       (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "BPI-63"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:232:                                     AAAATCGATTCTGTGGCTGG20                                                        (2) INFORMATION FOR SEQ ID NO:233:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 base pairs                                                     (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA                                                       (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "BPI-64"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:233:                                     AGAATCGATTTTGGTCATTA20                                                        (2) INFORMATION FOR SEQ ID NO:234:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 18 base pairs                                                     (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA                                                       (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "BPI-40"                                               (xi) SEQUENCE DESCRIPTION: SEQ ID NO:234:                                     TGATCTGAAGCTGGGCAG18                                                          (2) INFORMATION FOR SEQ ID NO:235:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 18 base pairs                                                     (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: DNA                                                       (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "BPI-7"                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:235:                                     GAACTTGGTTGTCAGTCG18                                                          (2) INFORMATION FOR SEQ ID NO:236:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 27 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "LBP Peptide 1-2"                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:236:                                     GlyGlnGlyLeuSerLeuSerIleSerAspSerSerIleArgValGln                              151015                                                                        GlyArgTrpLysValArgLysSerPhePheLys                                             2025                                                                          (2) INFORMATION FOR SEQ ID NO:237:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 22 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: misc.sub.-- feature                                             (D) OTHER INFORMATION: "LBP Peptide 2-1"                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:237:                                     AspValGluValAspMetSerGlyAspLeuGlyTrpLeuLeuAsnLeu                              151015                                                                        PheHisAsnGlnIleGlu                                                            20                                                                            __________________________________________________________________________

What is claimed are:
 1. In a method of treating a subject infected witha gram-positive bacterial species with an antibiotic, wherein thegram-positive bacterial species is selected from the group consisting ofBacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis,Staphylococcus hominis, Staphylococcus scirui, Staphylococcussaprophyticus, Staphylococcus haemolyticus, Staphylococcus hyicus,Staphylococcus intermedius, Staphylococcus simulans, Streptococcuspneumoniae, Streptococcus pyogenes, Streptococcus agalactia,Streptococcus bovis, Enterococcus faecalis, Enterococcus faecium,Enterococcus gallinarum, Enterococcus raffinosus, Enterococcuscasseliflavus and Enterococcus durans, the improvement comprising thestep of administering to said subject an amount of a BPI protein producteffective to increase the susceptibility of said gram-positive bacterialspecies to said antibiotic.
 2. The method of claim 1 wherein theantibiotic is selected from the group consisting of penicillins,cephalosporins, imipenem and monobactam antibiotics.
 3. The method ofclaim 1 wherein the antibiotic is an aminoglycoside antibiotic.
 4. Themethod of claim 1 wherein the antibiotic is a tetracycline antibiotic.5. The method of claim 1 wherein the antibiotic is a sulfonamide or atrimethoprim/sulfonamide antibiotic.
 6. The method of claim 1 whereinthe antibiotic is a fluoroquinolone antibiotic.
 7. The method of claim 1wherein the antibiotic is a macrolide antibiotic.
 8. The method of claim1 wherein the antibiotic is vancomycin.
 9. The method of claim 1 whereinthe antibiotic is selected from the group consisting of polymyxinantibiotics, chloramphenicol and lincosamide antibiotics.
 10. The methodof claim 1 wherein the gram-positive bacterial species is S. pneumoniaeand the antibiotic is a penicillin antibiotic.
 11. The method of claim 1wherein the gram-positive bacterial species is an Enterococcus speciesand the antibiotic is vancomycin.
 12. The method of claim 1 wherein thegram-positive bacterial species is a Staphylococcus species and theantibiotic is a penicillin antibiotic.
 13. In a method of treating asubject infected with a gram-positive bacterial species with anantibiotic, wherein the gram-positive bacterial species is resistant tosaid antibiotic and is selected from the group consisting of Bacillussubtilis, Staphylococcus aureus, Staphylococcus epidermidis,Staphylococcus hominis, Staphylococcus scirui, Staphylococcussaprophyticus, Staphylococcus haemolyticus, Staphylococcus hyicus,Staphylococcus intermedius, Staphylococcus simulans, Streptococcuspneumoniae, Streptococcus pyogenes, Streptococcus agalactia,Streptococcus bovis, Enterococcus faecalis, Enterococcus faecium,Enterococcus gallinarum, Enterococcus raffinosus, Enterococcuscasseliflavus and Enterococcus durans, the improvement comprising thestep of administering to said subject an amount of a BPI protein producteffective to reverse resistance of said gram-positive bacterial speciesto said antibiotic.
 14. The method of claim 13 wherein the antibiotic isselected from the group consisting of penicillins, cephalosporins,imipenem and monobactam antibiotics.
 15. The method of claim 13 whereinthe antibiotic is an aminoglycoside antibiotic.
 16. The method of claim13 wherein the antibiotic is a tetracycline antibiotic.
 17. The methodof claim 13 wherein the antibiotic is a sulfonamide or atrimethoprim/sulfonamide antibiotic.
 18. The method of claim 13 whereinthe antibiotic is a fluoroquinolone antibiotic.
 19. The method of claim13 wherein the antibiotic is a macrolide antibiotic.
 20. The method ofclaim 13 wherein the antibiotic is vancomycin.
 21. The method of claim13 wherein the antibiotic is selected from the group consisting ofpolymyxin antibiotics, chloramphenicol and lincosamide antibiotics. 22.The method of claim 13 wherein the gram-positive bacterial species is S.pneumoniae and the antibiotic is a penicillin antibiotic.
 23. The methodof claim 13 wherein the gram-positive bacterial species is anEnterococcus species and the antibiotic is vancomycin.
 24. The method ofclaim 13 wherein the gram-positive bacterial species is a Staphylococcusspecies and the antibiotic is a penicillin antibiotic.